TW201038734A - Polyunsaturated fatty acid synthase nucleic acid molecules and polypeptides, compositions, and methods of making and uses thereof - Google Patents

Polyunsaturated fatty acid synthase nucleic acid molecules and polypeptides, compositions, and methods of making and uses thereof Download PDF

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TW201038734A
TW201038734A TW099108150A TW99108150A TW201038734A TW 201038734 A TW201038734 A TW 201038734A TW 099108150 A TW099108150 A TW 099108150A TW 99108150 A TW99108150 A TW 99108150A TW 201038734 A TW201038734 A TW 201038734A
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polypeptide
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Kirk E Apt
Leslie Richter
David Simpson
Ross Zirkle
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Martek Biosciences Corp
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Abstract

The present invention is directed to isolated nucleic acid molecules and polypeptides of thraustochytrid polyunsaturated fatty acid (PUFA) synthases involved in the production of PUFAs, including PUFAs enriched in docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), or a combination thereof. The present invention is directed to vectors and host cells comprising the nucleic acid molecules, polypeptides encoded by the nucleic acid molecules, compositions comprising the nucleic acid molecules or polypeptides, and methods of making and uses thereof.

Description

201038734 六、發明說明： c考务明戶斤屬才支冬好々貝3 發明領域 本發明係針對涉及多不飽和脂肪酸(PUFA)的生產之多 不飽和脂肪酸(PUFA)合成酶的經單離的核酸分子及多肽， 包括富含二十二碳六烯酸(DHA)、二十碳五稀酸(EPA)，或 其等之一組合的PUFAs。本發明係針對包含該等核酸分子 的載體和宿主細胞、由該等核酸分子編碼的多肽、包含該 等核酸分子或多肽的組成物，以及製造其等之方法和其等 之用途。201038734 VI. INSTRUCTIONS: C. Examination of the genus of the genus of the polyunsaturated fatty acid (PUFA) synthetase involved in the production of polyunsaturated fatty acids (PUFA). Nucleic acid molecules and polypeptides, including PUFAs rich in docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), or a combination thereof. The present invention is directed to vectors and host cells comprising such nucleic acid molecules, polypeptides encoded by such nucleic acid molecules, compositions comprising such nucleic acid molecules or polypeptides, methods of making the same, and the like.

C先前#支J 發明背景 破囊菌為破囊壺菌目（Thraustochytriales)的微生 物，包括破囊壺菌屬與裂殖壺菌屬 少in’Mw)的成員，以及已經被承認為puFAs的一任 擇來源。參見，例如’美國專利案第5，130,242號。海洋細 菌與破囊壺菌中的聚酮合成酶(PKS)_類似系統近來已經顯 示出能夠自乙醯基-CoA與丙二醯基-CoA合成多不飽和脂 肪酸(PUFAs)。此等PKS合成酶-類似系統於本文中亦稱為 PUFA合成酶系統。海洋細菌内的puFA合成酶系統沙雷菌 (57^丽狀//α)與海洋弧菌（咖咖⑽)係於美國專利案第 6，140,486號之中說明。裂殖壺菌屬的破囊壺菌内的一puFA 合成酶系統係揭示於美國專利案第6,566,583號中。裂殖壺 菌屬與破囊壺菌屬的破囊壺菌内的PUFA合成酶系統亦於 3 201038734 美國專利案第7,247,461號之中說明。美國專利案第 7,211，418號說明破囊壺菌屬的破囊壺菌内的一？1^八合成 酶系統以及使用該系統生產的二十碳五烯酸(C20:5，亞米 加_3) (EPA)及其他的PUFAs。美國專利案第7,217,856號說 明沙雷菌〇S7zewane//a 〇"印〇77〇〇與日本沙雷菌 内的PUFA合成酶系統。WO 2005/097982說明菌株 SAM2179内的一PUFA合成酶系統。美國專利案第7,2〇8,590 號與第7,368,552號說明來自金黄色破囊壺菌 awrewm)的PUFA合成酶基因與蛋白質。 文獻中傳統上已經說明PKS系統屬於3種基本類型的 一者，典型地稱為第I型(模組式或迭代式）、第11型，以及第 III型。第I型模組式PKS系統也已經被稱為”模組式”？1(：8系 統，以及第I型迭代式PKS系統也已經被稱為”第1型，，1>1<：8系 統。第II型系統係特徵為可分離的蛋白質，其等之各個進行 有區別的酵素反應。該等酵雜力卫作以生產終產物，以 及該系 孤的合卿卿别的酵素典型地參與終產物的生產 次。此類型的系統係簡似於植物和細菌中發現的脂肪 合成酶(FA⑽制方式運作。第I型迭代式PKS系統與第 型系統係相似在於酵素係以迭代式的方式使用以生產终 物。第1型迭代式PKS系統與第II型系統不同在於較大的 白質之領域發生酵相活性，㈣是與可分離的蛋白質 關聯的。此系統細似於動物和真菌中發現的第職S 統0 與第II型系統相反， 第1型模組式p K S系統内的各酵素 201038734 領域於終產物的生產巾僅制—次。料領域係於非常大 的蛋白質内發現，以及各個反應的產物被傳遞至PKS蛋白 質内的另一個領域。 第III型系統已經更近來找到以及屬於縮合酵素之植物 的查耳酮(chalcone)合成酶家族。第III型PKS有區別於第工 型與第II型PKS系統以及於迭代式縮合反應中使用游離 CoA基質以通常產生一雜環系終產物。 於慣用的或標準的PUFA合成途徑中，中鏈長度飽和脂 肪酸（一脂肪酸合成酶(FAS)系統的產物）係藉由一系列的延 長和去飽和反應予以修飾。延長反應之基質為脂肪醯基 -CoA(要被延長的脂肪酸鏈）以及丙二醯基_c〇A(在各個延 長反應的期間所添加二個碳的來源）。延長酶反應的產物是 脂肪醯基-CoA，其具有二個額外的碳於直鏈中。去飽和酶 於業已存在的脂肪酸鏈中藉由於氧依賴性反應中取出二個 氳而創造順式雙鍵。去飽和酶的基質為醯基_C〇A(於一些動 物中）或者被酯化至磷脂質的甘油主幹(例如，磷脂醯膽鹼) 之脂肪酸。 脂肪酸係根據碳鏈的長度及飽和特徵來分類。脂肪酸 係根據鏈中的碳長度被稱為短鏈、中鏈或長鏈脂肪酸，當 碳原子之間沒有雙鍵存在時被稱為飽和脂肪酸，以及當雙 鍵存在時被稱為不飽和脂肪酸。當只有·一個雙鍵存在時， 不飽和長鏈脂肪酸為單不飽和的，以及當多於一個雙鍵存 在時，不飽和長键脂肪酸為多不飽和的。 PUFAs係根據離脂肪酸的甲基端之第一個雙鍵的位置 5 201038734 之來分類：亞米加-3(n-3)脂肪酸在第二個石厌上含有苐一個 雙鍵，而亞米加-6(η-6)脂肪酸在第六個奴上含有弟一個雙 鍵。舉例而言，二十二破六烯酸("DHA”)為具有22個碳的鏈 長及6個雙鍵的亞米加-3 PUFA ,通常稱為"22:6 η-3"。其他 的亞米加-3 PUFAs包括二十破五烯酸（”ΕΡΑ”）’稱為”20:5 η-3”，以及亞米加-3二十二碳五烯酸("DPAn-3’，）’稱為”22:5 n-3 ”。DHA與EPA已經被稱為「必須」脂肪酸。亞米加—6 PUFAs包括二十碳四烯酸("ARA")，稱為”20:4 n-6” ’以及亞 米加-6二十二碳五烯酸（"DPA n-6") ’稱為"22:5 n-6"。 亞米加-3脂肪酸為生物上重要的分子’其等由於其等 存在於細胞膜之中而影響細胞生理、調節生物活性化合物 的生產及基因表現，以及作用為生合成基質。Roche, Η. M., Proc. Nutr. Soc. 58: 397-401 (1999)。DHA(舉例而言，佔人 類大腦皮質内的脂質的大概15%-20%，以及佔視網膜内的 脂質的30%-60%)係集中於睪丸和精子，以及為母奶之重要 組份。Bergό,J.P.，andBamathan，G.Adv.Biochem·Eng·C Previously, the background of the genus Thraustochytriales is a member of the microorganisms of Thraustochytriales, including members of the genus Thraustochytrium and Schizochytrium, and has been recognized as a puFAs. Choose the source. See, for example, U.S. Patent No. 5,130,242. The polyketone synthase (PKS)-like system of marine bacteria and Thraustochytrium has recently been shown to be capable of synthesizing polyunsaturated fatty acids (PUFAs) from ethyl ketone-CoA and propylene glycol-CoA. Such PKS synthetase-similar systems are also referred to herein as PUFA synthase systems. The puFA synthase system in the marine bacteria, Serratia (57^丽状//α) and Marine Vibrio (Caf (10)) are described in U.S. Patent No. 6,140,486. A puFA synthase system in the genus Thraustochytrium of the genus Schizochytrium is disclosed in U.S. Patent No. 6,566,583. The PUFA synthase system in the genus Thraustochytrium and the Thraustochytrium is also described in U.S. Patent No. 7,247,461. US Patent No. 7,211,418 describes one of the thraustochytrid of the genus Thraustochytrium? 1^8 Synthetic enzyme system and eicosapentaenoic acid (C20:5, Amigas_3) (EPA) and other PUFAs produced using the system. U.S. Patent No. 7,217,856 describes the PUFA synthase system of Serratia serrata S7zewane//a 〇"Indigo 77〇〇 and Serratia marcescens. WO 2005/097982 describes a PUFA synthase system within strain SAM2179. U.S. Patent Nos. 7,2,8,590 and 7,368,552 describe PUFA synthase genes and proteins from Thraustochytrium awrewm. The PKS system has traditionally been described in the literature as belonging to one of three basic types, typically referred to as Type I (modular or iterative), Type 11, and Type III. The Type I modular PKS system has also been called "modular"? The 1(:8 system, and the Type I iterative PKS system has also been referred to as the "Type 1," > 1 <:8 system. The Type II system is characterized by separable proteins, and so on. There are distinct enzyme reactions. These enzymes are used to produce the final product, and the enzymes of the genus of the genus are typically involved in the production of the final product. This type of system is similar to plants and bacteria. The discovered fat synthase (FA(10) system works. The type I iterative PKS system is similar to the first system in that the enzyme is used in an iterative manner to produce the final product. Type 1 iterative PKS system and type II system The difference lies in the activity of the yeast phase in the field of larger white matter, and (iv) the association with the separable protein. This system is similar to that found in animals and fungi. The first S system is the opposite of the type II system. Each of the enzymes in the group p KS system 201038734 is produced only in the production of the final product. The material field is found in very large proteins, and the products of each reaction are transferred to another field in the PKS protein. Type III The chalcone synthase family of plants belonging to the condensed enzymes has been more recently found. Type III PKS differs from the first type and the second type PKS system and uses a free CoA matrix in an iterative condensation reaction to A heterocyclic end product is produced. In a conventional or standard PUFA synthesis pathway, medium chain length saturated fatty acids (products of a fatty acid synthase (FAS) system) are modified by a series of extension and desaturation reactions. The substrate for prolonging the reaction is fat thiol-CoA (the fatty acid chain to be extended) and propylenediyl-c〇A (the source of two carbons added during each extended reaction). The product of the extended enzymatic reaction is fat. Mercapto-CoA, which has two additional carbons in a linear chain. The desaturase creates a cis-double bond in the existing fatty acid chain by taking two deuteriums in an oxygen-dependent reaction. It is a fatty acid of thiol_C〇A (in some animals) or a glycerol backbone (eg, phospholipid choline) esterified to a phospholipid. Fatty acids are classified according to the length and saturation characteristics of the carbon chain. Fatty acids are called short-chain, medium-chain or long-chain fatty acids according to the length of carbon in the chain. When there is no double bond between carbon atoms, they are called saturated fatty acids, and when double bonds are present, they are called unsaturated fatty acids. Unsaturated long-chain fatty acids are monounsaturated when only one double bond is present, and unsaturated long-chain fatty acids are polyunsaturated when more than one double bond is present. PUFAs are based on methyl groups derived from fatty acids. The position of the first double bond on the end 5 201038734 Classification: Amiga-3 (n-3) fatty acid contains a double bond on the second stone, and Amiga-6 (η-6) Fatty acid contains a double bond in the sixth slave. For example, docosahexaenoic acid ("DHA" is an Amiga-3 PUFA with a chain length of 22 carbons and 6 double bonds. , usually called "22:6 η-3". Other Amiga-3 PUFAs include T-pentaenoic acid ("ΕΡΑ")' called "20:5 η-3", and Amiga-3 docosapentaenoic acid ("DPAn-3 ',) ' is called "22:5 n-3". DHA and EPA have been called "essential" fatty acids. Amiga-6 PUFAs include arachidonic acid ("ARA"), called "20:4 n-6" and Amiga -6 docosapentaenoic acid ("DPA n- 6") 'called "22:5 n-6". The sub-male-3 fatty acid is a biologically important molecule which affects cell physiology, regulates the production of biologically active compounds and gene expression, and acts as a biosynthetic matrix due to its presence in the cell membrane. Roche, Η. M., Proc. Nutr. Soc. 58: 397-401 (1999). DHA (for example, about 15%-20% of lipids in the human cerebral cortex and 30%-60% of lipids in the retina) are concentrated in testis and sperm, and are important components of breast milk. Bergό, J.P., andBamathan, G.Adv.Biochem·Eng·

Biotechnol· 96:49-125 (2005)。DHA佔腦内之亞米加-3脂肪 酸的多至97°/。以及佔視網膜内之亞米加_3脂肪酸的多至 93。/。。而且’ DHA對胎兒和嬰兒的發展為不可缺的，以及 成人之認知功能的維持。同前。因為亞米加_3脂肪酸於人 類身體内不會被重新合成，此等脂肪酸必須自營養的來源 衍生。 亞麻子油和魚油被認為是亞米加_3脂肪酸之良好的飲 食來源。亞麻子油不含有EPA、DHA、DpA，或是ARA， 201038734 而是含有亞麻油酸(C18:3 η-3)，使身體能製造EPA之建造區 塊。然而，有證據顯示代謝轉化的速率可以是缓慢且玎變 的，尤其於健康受損的該等之中。脂肪酸組成物的類蜇和 位準中之魚油有相當地變化，取決於特定的物種以及其等 之飲食。舉例而言，由水產養殖倒養的魚傾向比野生的該 等魚具有較低位準的亞米加-3脂肪酸。再者，魚油帶有含 有環境汙染的風險，以及可以與安定性問題及魚的氣味或 味道相關聯。 由破囊壺菌生產的油通常具有比對應的魚或微藻類油 更間卓的多不飽和脂肪酸形態。Lewis，T.E.，Mar. Biotechnol. 1: 580-587 (1999)。破囊壺菌物種之菌株已被報 導產生亞米加-3脂肪酸為該等有機體生產之高百分比的總 脂肪酸。美國專利案第5，130,242號；Huang，J.等人，J. Am. Oil. Chem. Soc. 78: 605-610 (2001) ; Huang, J.等人，Mar.Biotechnol 96: 49-125 (2005). DHA accounts for up to 97°/ of the sub-male-3 fatty acid in the brain. And up to 93% of the amino acid _3 fatty acids in the retina. /. . Moreover, 'DHA is essential for the development of fetuses and babies, as well as the maintenance of cognitive function in adults. Cit. Because the amiga _3 fatty acids are not re-synthesized in the human body, these fatty acids must be derived from the source of nutrition. Linseed oil and fish oil are considered to be good sources of food for Yamiga _3 fatty acids. Linseed oil does not contain EPA, DHA, DpA, or ARA, 201038734 but contains linoleic acid (C18:3 η-3), which allows the body to manufacture EPA building blocks. However, there is evidence that the rate of metabolic conversion can be slow and mutated, especially among those with impaired health. The fatty acids and the fish oil in the grades vary considerably depending on the particular species and its diet. For example, fish reared by aquaculture tend to have lower levels of sub-male-3 fatty acids than wild ones. Furthermore, fish oil carries a risk of environmental pollution and can be associated with stability issues and the smell or taste of the fish. Oils produced by Thraustochytrium usually have a more polyunsaturated fatty acid form than the corresponding fish or microalgae oil. Lewis, T.E., Mar. Biotechnol. 1: 580-587 (1999). Strains of Thraustochytrium species have been reported to produce a high percentage of total fatty acids produced by the Amiga-3 fatty acids. U.S. Patent No. 5,130,242; Huang, J. et al., J. Am. Oil. Chem. Soc. 78: 605-610 (2001); Huang, J. et al., Mar.

Biotechnol. 5: 450_457 (2〇03)。然而，經單離的破囊壺菌於 生產的PUFAs之同一性及量上有變化，藉此一些先前說明 的菌株能具有非所欲的PUFA形態。 已經努力藉由内源性產生的脂肪酸之修飾來生產油杆 作物植物内之PUFAs。用脂肪酸延長酶和去飽和酶之各種 各樣的個別的基因之此等植物之基因修飾已經生產出葉子 或種子’其等含有可測量的位準之PUFAs(例如：EPA)，但 是也含有顯著位準之混合的更短鏈的和較不飽和的 PUFAs(Qi等人，Nature Biotech. 22:739 (2004); PCT PublBiotechnol. 5: 450_457 (2〇03). However, the isolated colony of Thraustochytrium has a change in the identity and amount of PUFAs produced, whereby some of the previously described strains can have an undesired PUFA morphology. Efforts have been made to produce PUFAs in oilseed crop plants by modification of endogenously produced fatty acids. Genetic modification of such plants with various individual genes of fatty acid elongases and desaturases has produced leaves or seeds that have measurable levels of PUFAs (eg, EPA), but also contain significant Mixed shorter chain and less saturated PUFAs (Qi et al, Nature Biotech. 22:739 (2004); PCT Publ

No. WO 04/071467 ; Abbadi等人，Plant Cell 16:1 (2〇〇4)); 7 201038734No. WO 04/071467; Abbadi et al., Plant Cell 16:1 (2〇〇4)); 7 201038734

Napier and Sayanova, Proc. Nutrition Society 64:387-393 (2005) ; Robert等人，Functional Plant Biology 32:473-479 (2005);以及美國申請公開案第2004/0172682號）。 確切而言，對於與所欲的PUFA形態關聯的核酸分子及 多狀之早離以及經由使用此等核酸分子及多狀來產生所欲 的PUFA形態的方法存在持續性的需求。 L發明内容3 發明概要 本發明係針對一經單離的核酸分子，其係選自於以下 所構成的群組：0)—核酸分子，其包含與序列辨識編號：1 有至少80%的同一性之多核苷酸序列，其中該多核苷酸序 列編碼一多肽，其包含選自於以下所構成的群組之PUFA合 成酶活性：β-酮基醯基-ACP合成酶(KS)活性、丙二醯基 -CoA : ACP醢基轉移酶(MAT)活性、醯基載體蛋白質（ACP) 活性、酮還原酶(KR)活性、β-羥基醯基-ACP脫水酶(DH)活 性，以及其等之組合；（b)—核酸分子，其包含與序列辨識 編號：7有至少80%的同一性之多核苷酸序列，其中該多核 苷酸序列編碼一多肽’其包含KS活性；（c)一核酸分子，其 包含與序列辨識編號.9有至少80¾的同—'性之多核普酸序 列，其中該多核苷酸序列編碼一多肽，其包含MAT活性； (d) —核酸分子’其包含與序列辨識編號：13、15、17、19、 21，或23的任一者有至少80%的同一性之多核苷酸序列， 其中該多核苷酸序列編碼一多肽，其包含ACP活性；（e) — 核酸分子，其包含與序列辨識編號：11有至少80%的同一 201038734 性之多核苷酸序列，其中該多核苷酸序列編碼一多肽，其 包含ACP活性；（f)一核酸分子，其包含與序列辨識編號： 25有至少80%的同一性之多核苷酸序列，其中該多核苷酸 序列編碼一多肽，其包含KR活性；以及(g)—核酸分子，其 包含與序列辨識編號：27有至少80%的同一性之多核苷酸 序列，其中該多核苷酸序列編碼一多肽，其包含DH活性。 於一些具體例中，該多核苷酸序列係與序列辨識編號：1、 7、9、11、13、15、17、19、21、23、25，及 27 有至少 90% 〇 的同一性或有至少95%的同一性。於一些具體例中，該等 核酸分子各別包含序列辨識編號：1、7、9、11、13、15、 17、19、21、23、25，及27之多核苷酸序列。 本發明係針對一經單離的核酸分子，其係選自於以下 所構成的群組：（a)—核酸分子，其包含編碼一多肽之多核 苷酸序列，其中該多肽包含與序列辨識編號：2有至少80% 的同一性之胺基酸序列，以及其中該多肽包含選自於以下 ^ 所構成的群組之PUFA合成酶活性：KS活性、MAT活性、Napier and Sayanova, Proc. Nutrition Society 64: 387-393 (2005); Robert et al, Functional Plant Biology 32: 473-479 (2005); and U.S. Application Publication No. 2004/0172682). Specifically, there is a continuing need for methods for generating the desired PUFA morphology for nucleic acid molecules associated with the desired PUFA morphology and for early isolation and for the use of such nucleic acid molecules and polymorphisms. SUMMARY OF THE INVENTION The present invention is directed to an isolated nucleic acid molecule selected from the group consisting of: 0) a nucleic acid molecule comprising at least 80% identity to the sequence identification number: A polynucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising PUFA synthase activity selected from the group consisting of: β-ketothiol-ACP synthase (KS) activity, C Dimercapto-CoA: ACP thiol transferase (MAT) activity, thiol-based carrier protein (ACP) activity, ketoreductase (KR) activity, β-hydroxy thiol-ACP dehydratase (DH) activity, and the like a combination of (b) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 7, wherein the polynucleotide sequence encodes a polypeptide comprising KS activity; (c) A nucleic acid molecule comprising at least 803 of the same-sex multi-nucleotide sequence as sequence identification number .9, wherein the polynucleotide sequence encodes a polypeptide comprising MAT activity; (d) - a nucleic acid molecule Contains any of the serial identification numbers: 13, 15, 17, 19, 21, or 23 a polynucleotide sequence having at least 80% identity, wherein the polynucleotide sequence encodes a polypeptide comprising ACP activity; (e) a nucleic acid molecule comprising at least 80% identical to the sequence ID: 11 201038734 A polynucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising ACP activity; (f) a nucleic acid molecule comprising a polynucleotide having at least 80% identity to sequence identification number: 25. a sequence, wherein the polynucleotide sequence encodes a polypeptide comprising KR activity; and (g) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 27, wherein the polynucleotide The nucleoside sequence encodes a polypeptide comprising DH activity. In some embodiments, the polynucleotide sequence has at least 90% 〇 identity with sequence identification numbers: 1, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, and 27. Have at least 95% identity. In some embodiments, the nucleic acid molecules each comprise a polynucleotide sequence of sequence identification numbers: 1, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, and 27. The present invention is directed to an isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises a sequence identification number : 2 an amino acid sequence having at least 80% identity, and wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of KS activity, MAT activity,

U ACP活性、KR活性、DH活性、以及其等之組合；（b)—核 酸分子，其包含編碼一多肽之多核苷酸序列，其中該多肽 包含與序列辨識編號：8有至少80%的同一性之胺基酸序 列，以及其中該多肽包含KS活性；（c)一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：10有至少80%的同一性之胺基酸序列，以及其中該 多肽包含MAT活性；（d)—核酸分子，其包含編碼一多肽之 多核苷酸序列，其中該多肽包含與序列辨識編號：14、16、 9 201038734 18、20、” ~v、u Z ’或24的任何一者有至少80%的同一性之胺基 西文序列，以及其中1亥多肽包含ACP活性；（e)-核酸分子’ 八13編碼—多肽之多核苷酸序列，其中該多肽包含與序 列辨識編號：12有至少8()%的同—性之胺基酸序列，以及 ”中"亥夕肽包含ACP活性；⑴一核酸分子，其包含編碼一 夕肽之多核苷酸序列’其中該多肽包含與序列辨識編號： %有至少8〇%的同—性之胺基酸序列以及其中該多肽包 a KR活性，以及(g) —核酸分子其包含編碼一多肽之多核 苷酸序列，其中該多肽包含與序列辨識編號：28有至少8〇% 的同—性之胺基酸序列，以及其中該多肽包含DH活性。於 些具體例中’該胺基酸序列係各別與序列辨識編號：2、 8、10、1〇 ^、14、16、18、20、22、24、26，及28有至少 90% 的同性或有至少95。/〇的同一性。於一些具體例中，該等 多狀各別包含序列辨識編號：2 ' 8、10、12、14、16、18、 ' 22 ' 24、26，及28之胺基酸序列。 本發明係針對一經單離的核酸分子，其係選自於以下 所構成的群組：（a)—核酸分子，其包含與序列辨識編號：3 有至少80%的同一性之多核苷酸序列，其中該多核苷酸序 列編碼一多肽，其包含選自於以下所構成的群組之PUFA合 成酶活性：KS活性、鏈長度因子(CLF)活性、醯基轉移酶(AT) 活性、烯醯基-ACP還原酶(enoyl-ACPreductase)(ER)活性， 以及其等之組合；（b)一核酸分子’其包含與序列辨識編號： 29有至少80%的同一性之多核苷酸序列，其中該多核苷酸 序列編碼一多肽’其包含KS活性；（c)一核酸分子’其包含 201038734 與序列辨識編號：31有至少80%的同一性之多核苷酸序 列，其中該多核苷酸序列編碼一多肽，其包含CLF活性；（d) 一核酸分子，其包含與序列辨識編號：33有至少80%的同 一性之多核苷酸序列，其中該多核苷酸序列編碼一多肽， 其包含AT活性；以及(e)—核酸分子，其包含與序列辨識編 號：35有至少80%的同一性之多核苷酸序列，其中該多核 苷酸序列編碼一多肽，其包含ER活性。於一些具體例中， 該多核苷酸序列係各別與序列辨識編號：3、29、31、33， 〇 以及35有至少90%的同一性或有至少95%的同一性。於一些 具體例中，該等核酸分子各別包含序列辨識編號：3、29、 31、33，以及35的多核苷酸序列。 本發明係針對一經單離的核酸分子，其係選自於以下 所構成的群組：（a)—核酸分子，其包含編碼一多肽之多核 苷酸序列，其中該多肽包含與序列辨識編號：4有至少80% 的同一性之胺基酸序列，以及其中該多肽包含選自於以下a combination of U ACP activity, KR activity, DH activity, and the like; (b) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% of the sequence identification number: An amino acid sequence of identity, and wherein the polypeptide comprises KS activity; (c) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% of the sequence identification number: An amino acid sequence of identity, and wherein the polypeptide comprises MAT activity; (d) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises and the sequence identification number: 14, 16, 9 201038734 18, 20, " any of ~v, u Z ' or 24 has at least 80% identity of the amino-Western sequence, and wherein the 1 HI polypeptide comprises ACP activity; (e)-nucleic acid molecule '8-13 coding a polynucleotide sequence of a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 8 (%) homology to the sequence number: 12, and "Chinese" has a ACP activity; (1) a nucleic acid molecule Containing the peptide a nucleotide sequence 'wherein the polypeptide comprises a homologous amino acid sequence having a sequence identification number: % of at least 8 % and wherein the polypeptide comprises a KR activity, and (g) - the nucleic acid molecule comprises a coding number A polynucleotide sequence of a peptide, wherein the polypeptide comprises at least 8% homozygous amino acid sequence with sequence number: 28, and wherein the polypeptide comprises DH activity. In some embodiments, the amino acid sequence is at least 90% different from the sequence identification numbers: 2, 8, 10, 1〇, 14, 16, 18, 20, 22, 24, 26, and 28. Same sex or have at least 95. /〇 The identity. In some embodiments, the plurality of sequences each comprise a sequence identification number: 2' 8, 10, 12, 14, 16, 18, '22' 24, 26, and 28 amino acid sequences. The present invention is directed to an isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: Wherein the polynucleotide sequence encodes a polypeptide comprising PUFA synthase activity selected from the group consisting of KS activity, chain length factor (CLF) activity, thiol transferase (AT) activity, olefin a thiol-ACP reductase (ER) activity, and combinations thereof; (b) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 29, Wherein the polynucleotide sequence encodes a polypeptide comprising KS activity; (c) a nucleic acid molecule comprising a polynucleotide sequence of 201038734 having at least 80% identity to sequence identification number: 31, wherein the polynucleotide The sequence encodes a polypeptide comprising CLF activity; (d) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to Sequence ID: 33, wherein the polynucleotide sequence encodes a polypeptide, It contains AT activity; and (e) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to the sequence recognition number: 35, wherein the polynucleotide sequence encodes a polypeptide comprising ER activity. In some embodiments, the polynucleotide sequences are each at least 90% identical or at least 95% identical to the sequence identification numbers: 3, 29, 31, 33, 〇 and 35. In some embodiments, the nucleic acid molecules each comprise a polynucleotide sequence of sequence identification numbers: 3, 29, 31, 33, and 35. The present invention is directed to an isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises a sequence identification number : 4 having at least 80% identity amino acid sequence, and wherein the polypeptide comprises selected from the group consisting of

q 所構成的群組之PUFA合成酶活性：KS活性、CLF活性、AT 活性、ER活性、以及其等之組合；（b)—核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：30有至少80%的同一性之胺基酸序列，以及其中該 多肽包含KS活性；（c)一核酸分子，其包含編碼一多肽之多 核苷酸序列，其中該多肽包含與序列辨識編號：32有至少 80%的同一性之胺基酸序列，以及其中該多肽包含CLF活 性；（d)—核酸分子，其包含編碼一多肽之多核苷酸序列， 其中該多肽包含與序列辨識編號：34有至少80%的同一性 11 201038734 之胺基酸序列，以及其中該多肽包含AT活性；以及(e)—核 酸分子，其包含編碼一多肽之多核苷酸序列，其中該多肽 包含與序列辨識編號：36有至少80%的同一性之胺基酸序 列，以及其中該多肽包含ER活性。於一些具體例中，該胺 基酸序列係各別與序列辨識編號：4、30、32、34，及36有 至少90%的同一性或有至少95%的同一性。於一些具體例 中，該多肽各別包含序列辨識編號：4、30、32、34，及36 的胺基酸序列。 本發明係針對一經單離的核酸分子，其係選自於以下 所構成的群組：0)—核酸分子，其包含與序列辨識編號：5 有至少80%的同一性之多核苷酸序列，其中該多核苷酸序 列編碼一多肽，其包含選自於以下所構成的群組之PUFA合 成酶活性：DH活性、ER活性、以及其等之組合；（b)—核 酸分子，其包含與序列辨識編號：37有至少80%的同一性 之多核苷酸序列，其中該多核苷酸序列編碼一多肽，其包 含DH活性；（c)一核酸分子，其包含與序列辨識編號：39 有至少80%的同一性之多核苷酸序列，其中該多核苷酸序 列編碼一多肽，其包含DH活性；以及(d)—核酸分子，其包 含與序列辨識編號：41有至少80%的同一性之多核苷酸序 列，其中該多核苷酸序列編碼一多肽，其包含ER活性。於 一些具體例中，該多核苷酸序列係各別與序列辨識編號： 5、37、39，及41有至少90%的同一性或有至少95°/。的同一 性。於一些具體例中，該等核酸分子各別包含序列辨識編 號：5、37、39，及41的多核苷酸序列。 12 201038734 本發明係針對一經單離的核酸分子，其係選自於以下 所構成的群組：（a)—核酸分子，其包含編碼一多肽之多核 苷酸序列，其中該多肽包含與序列辨識編號：6有至少80% 的同一性之胺基酸序列，以及其中該多肽包含選自於以下 所構成的群組之PUFA合成酶活性：DH活性、ER活性、以 及其等之組合；（b)—核酸分子，其包含編碼一多肽之多核 苷酸序列，其中該多肽包含與序列辨識編號：38有至少80% 的同一性之胺基酸序列，以及其中該多肽包含DH活性；（c) 一核酸分子，其包含編碼一多肽之多核苷酸序列，其中該 多肽包含與序列辨識編號：40有至少80%的同一性之胺基 酸序列，以及其中該多肽包含DH活性；以及(d)—核酸分 子，其包含編碼一多肽之多核苷酸序列，其中該多肽包含 與序列辨識編號：42有至少80%的同一性之胺基酸序列， 以及其中該多肽包含ER活性。於一些具體例中，該胺基酸 序列係各別與序列辨識編號：6、38、40，以及42有至少90% 的同一性或有至少的95%同一性。於一些具體例中，該等 多肽各別包含序列辨識編號：6、38、40，以及42之胺基酸 序列。 本發明係針對一經單離的核酸分子，其係選自於以下 所構成的群組：（a)—核酸分子，其包含與序列辨識編號： 68或序列辨識編號：120有至少80%的同一性之多核苷酸序 列，其中該多核苷酸序列編碼一多肽，其包含選自於以下 所構成的群組之PUFA合成酶活性：KS活性、MAT活性、 ACP活性、KR活性、DH活性、以及其等之組合；（b)—核 13 201038734 酸分子，其包含與序列辨識編號：74有至少80%的同一性 之多核苷酸序列，其中該多核苷酸序列編碼一多肽，其包 含KS活性；（c)一核酸分子，其包含與序列辨識編號：76有 至少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含MAT活性；（d)—核酸分子，其包含與 序列辨識編號：80、82、84、86、88、90、92、94、96或 98的任一者有至少80%的同一性之多核苷酸序列，其中該 多核苷酸序列編碼一多肽，其包含ACP活性；（e) —核酸分 子，其包含與序列辨識編號：78有至少80%的同一性之多 核苷酸序列，其中該多核苷酸序列編碼一多肽，其包含ACP 活性；（f)一核酸分子，其包含與序列辨識編號：100有至少 80%的同一性之多核苷酸序列，其中該多核苷酸序列編碼 一多肽，其包含KR活性；以及(g)—核酸分子，其包含與序 列辨識編號：118有至少80%的同一性之多核苷酸序列，其 中該多核苷酸序列編碼一多肽，其包含DH活性。於一些具 體例中，該多核苷酸序列係各別與序列辨識編號：68、74、 76、78、80、82、84、86、88、90、92、94、96、98、100、 118，及120有至少90°/。的同一性或有至少95%的同一性。於 一些具體例中，該等核酸分子各別包含序列辨識編號：68、 74、76、78、80、82、84、86、88、90、92、94、96、98、 100、118，及120的多核苷酸序列。 本發明係針對一經單離的核酸分子，其係選自於以下 所構成的群組：（a)—核酸分子，其包含編碼一多肽之多核 苷酸序列，其中該多肽包含與序列辨識編號：69有至少80% 14 201038734 的同一性之胺基酸序列，以及其中該多肽包含選自於以下 所構成的群組之PUFA合成酶活性：KS活性、MAT活性、 ACP活性、KR活性、DH活性、以及其等之組合；（b)—核 酸分子，其包含編碼一多肽之多核苷酸序列，其中該多肽 包含與序列辨識編號：75有至少80%的同一性之胺基酸序 列，以及其中該多肽包含KS活性；（c)一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：77有至少80%的同一性之胺基酸序列，以及其中該 〇 多肽包含MAT活性；（d)—核酸分子，其包含編碼一多肽之 多核苷酸序列，其中該多肽包含與序列辨識編號：81、83、 85、87、89、91、93、95、97或99 的任一者有至少 80%的 同一性之胺基酸序列，以及其中該多肽包含ACP活性；（e) 一核酸分子，其包含編碼一多肽之多核苷酸序列，其中該 多肽包含與序列辨識編號：79有至少80%的同一性之胺基 酸序列，以及其中該多肽包含ACP活性；（f)一核酸分子， Q 其包含編碼一多肽之多核苷酸序列，其中該多肽包含與序 列辨識編號：101有至少80%的同一性之胺基酸序列，以及 其中該多肽包含KR活性；以及(g)—核酸分子，其包含編碼 一多肽之多核苷酸序列，其中該多肽包含與序列辨識編 號：119有至少80%的同一性之胺基酸序列，以及其中該多 肽包含DH活性。於一些具體例中，該胺基酸序列係各別與 序歹㈣識編號：69、75、77、79、81、83、85、87、89、 91、93、95、97、99、101，以及119有至少 90%的同一性 或有至少95%的同一性。於一些具體例中，該等多肽各別 15 201038734 包含序列辨識編號：69、75、77、79、81、83、85、87、 89、91、93、95、97、99、101，以及 119之胺基酸序列。 本發明係針對一經單離的核酸分子，其係選自於以下 所構成的群組：（a)—核酸分子’其包含與序列辨識編號： 70或序列辨識編號：121有至少80%的同一性之多核苷酸序 列，其中該多核苷酸序列編碼一多肽，其包含選自於以下 所構成的群組之PUFA合成酶活性：KS活性、鏈長度因子 (CLF)活性、醯基轉移酶(AT)活性、烯醯基-ACP還原酶(ER) 活性，以及其等之組合；（b)—核酸分子，其包含與序列辨 識編號：102有至少80%的同一性之多核苷酸序列，其中該 多核苷酸序列編碼一多肽，其包含KS活性；（c)一核酸分 子，其包含與序列辨識編號：1〇4有至少80%的同一性之多 核苦酸序列，其中該多核音酸序列編碼一多狀，其包含CLF 活性；（d)—核酸分子，其包含與序列辨識編號：1〇6有至少 80%的同一性之多核苷酸序列’其中該多核苷酸序列編碼 一多肽，其包含AT活性，以及(e)—核酸分子，其包含與序 列辨識編號：108有至少80%的同一性之多核苷酸序列，其 中該多核苷酸序列編碼一多肽，其包含ER活性。於一些具 體例中，該多核苷酸序列係各別與序列辨識編號：7〇、1〇2、 104、106、108，以及121有至少90°/。的同一性或有至少95% 的同一性。於一些具體例中，該等核酸分子各別包含序列 辨識編號：70、102、104、1〇6、108,以及121的多核苦酸 序列。 本發明係針對一經單離的核酸分子，其係選自於以下 16 201038734 所構成的群組：（a)一核酸分子，其包含編碼一多肽之多核 苦酸序列，其中該多肽包含與序列辨識編號：71有至少80% 的同一性之胺基酸序列，以及其中該多肽包含選自於以下 所構成的群組之PUFA合成酶活性：KS活性、CLF活性、AT 活性、ER活性 '以及其等之組合；（b)—核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：103有至少8〇%的同一性之胺基酸序列，以及其中該 〇 多狀包含KS活性；（c)一核酸分子，其包含編碼一多肽之多 核普酸序列’其中該多肽包含與序列辨識編號：105有至少 8〇%的同—性之胺基酸序列，以及其中該多肽包含CLF活 • 性’（d)—核酸分子，其包含編碼一多肽之多核苷酸序列， . 其中S亥多肽包含與序列辨識編號：107有至少80%的同一性 之胺基酸序列，以及其中該多肽包含Ατ活性；以及一核 酉文刀子，其包含編碼一多肽之多核苷酸序列，其中該多肽 包含與序列辨識編號：109有至少80%的同一性之胺基酸序 Ο 歹·！以及其中s亥多狀包含ER活性。於一些具體例中，該胺 基酸序列係各別與序列辨識編號：71、1〇3、1〇5、1〇7，以 及109有至少9〇%的同一性或有至少％%的同一性。於一些 具體例中，該多肽各別包含序列辨識編號：71、1〇3、1〇5、 107，以及1〇9的胺基酸序列。 本發明係針對一經單離的核酸分子，其係選自於以下 所構成的群組：（a)一核酸分子，其包含與序列辨識編號： 72或序列辨識編號：122有至少8〇%的同一性之多核苷酸序 列，其中該多核苷酸序列編碼一多肽，其包含選自於以下 17 201038734 所構成的群組之PUFA合成酶活性：DH活性、ER活性、以 及其等之組合；（b)—核酸分子，其包含與序列辨識編號： 110有至少80%的同一性之多核苷酸序列，其中該多核苷酸 序列編碼一多肽，其包含DH活性；（c) 一核酸分子，其包含 與序列辨識編號：112有至少80%的同一性之多核苷酸序 列，其中該多核苷酸序列編碼一多肽，其包含DH活性；以 及(d)—核酸分子，其包含與序列辨識編號：114有至少80% 的同一性之多核苷酸序列，其中該多核苷酸序列編碼一多 肽，其包含ER活性。於一些具體例中，該多核苷酸序列係 各別與序列辨識編號：72、110、112、114，以及122有至 少90%的同一性或有至少95%的同一性。於一些具體例中， 該等核酸分子各別包含序列辨識編號：72、110、112、114， 以及122的該等多核苷酸序列。 本發明係針對一經單離的核酸分子，其係選自於以下 所構成的群組：（a)—核酸分子，其包含編碼一多肽之多核 苷酸序列，其中該多肽包含與序列辨識編號：73有至少80% 的同一性之胺基酸序列，以及其中該多肽包含選自於以下 所構成的群組之PUFA合成酶活性：DH活性、ER活性、以 及其等之組合；（b)—核酸分子，其包含編碼一多肽之多核 苷酸序列，其中該多肽包含與序列辨識編號：111有至少80% 的同一性之胺基酸序列，以及其中該多肽包含DH活性；（c) 一核酸分子，其包含編碼一多肽之多核苷酸序列，其中該 多肽包含與序列辨識編號：113有至少80%的同一性之胺基 酸序列，以及其中該多肽包含DH活性；以及（d) —核酸分 18 201038734 子，其包含編碼一多肽之多核苷酸序列，其中該多肽包含 與序列辨識編號：115有至少80%的同一性之胺基酸序列， 以及其中該多肽包含ER活性。於一些具體例中，該胺基酸 序列係各別與序列辨識編號：73、111、113，以及115有至 少90%的同一性或有至少95%的同一性。於一些具體例中， 該等多肽各別包含序列辨識編號：73、111、113，以及115 之胺基酸序列。 本發明係針對一種經單離的核酸分子，其包含編碼一 多肽之多核苷酸序列，該多肽包含選自於以下所構成的群 組之PUFA合成酶活性：KS活性、MAT活性、ACP活性、 KR活性、CLF活性、AT活性、ER活性、DH活性、以及其 等之組合，其中該多核苷酸係於嚴苛條件下與以上說明的 該等多核苷酸序列的任一者之該互補物雜交。 本發明係針對一種經單離的核酸分子，其包含一多核 苷酸序列，其係與以上說明的該等多核苷酸序列的任一者 完全互補的。 本發明係針對一種重組型核酸分子，其包含：以上說 明的該等核酸分子的任一者或其等之組合以及一轉錄控制 序列。於一些具體例中，該重組型核酸分子為一重組型載 體。 本發明係針對一宿主細胞，其表現以上說明的該等核 酸分子的任一者、以上說明的該等重組型核酸分子的任一 者，以及其等之組合。於一些具體例中，該宿主細胞係選 自於以下所構成的群組：植物細胞、微生物細胞，以及動 19 201038734 物細胞。於一些具體例中，微生物細胞為一細菌。於一些 具體例中，該細菌為大腸桿菌。於一些具體例中，該細菌 為海洋的細菌。於一些具體例中’微生物細胞為破囊壺菌。 於一些具體例中，該破囊壺菌係為裂殖壺菌。於一些具體 例中，該破囊壺菌係為破囊壺菌。於一些具體例中，該破 囊壺菌為巫肯尼亞菌（。 本發明係針對一種產生至少一 PUFA的方法，其包含： 於有效產生PUFA的條件下於一宿主細胞中表現PUFA合成 酶基因，其中該PUFA合成酶基因包含以上說明的該等經單 離的核酸分子的任一者、以上說明的該等重組型核酸分子 的任一者，或其等之組合，以及其中至少一PUFA係被生 產。於本具體例的一態樣中，該宿主細胞係選自於以下所 構成的群組：植物細胞、經單離的動物細胞，以及微生物 細胞。於本具體例的另一態樣中，該至少一個PUFA包含二 十二碳六烯酸(DHA)或二十碳五烯酸(EPA)。 本發明係針對一種生產富含DHA、EPA，或其等之一 組合的脂質的方法，其包含：於有效產生脂質的條件下於 伤主細胞中表現pUFA合成酶基因，其中該PUFA合成酶 基因包含以上說明的該等經單離的核酸分子的任一者、以 上說明的該等重組型核酸分子的任一者，或其等之組合於 "亥佰主細胞中，以及其中富含DHA、，或其等之一組 σ的脂質係被生產。本發明係針對一種製造一重組型載體 的方法，其包含將以上說明的該等經單離的核酸分子的任 —者插入至載體内。 20 201038734 本發明係針對一種製造重組型宿主細胞的方法，其包 含將一重組型載體引入如以上說明的之至一宿主細胞中。 於一些具體例中，該宿主細胞係選自於以下所構成的群 組：植物細胞、經單離的動物細胞，以及微生物細胞。 本發明係針對由所編碼之經單離的多肽以上說明的該 等多核苷酸序列的任一者。 本發明係針對一種經單離的多肽，其係選自於以下所 構成的群組：（a)—多肽，其包含與序列辨識編號：2有至少 80%的同一性之胺基酸序列，其中該多肽包含選自於以下 所構成的群組之PUFA合成酶活性：KS活性、MAT活性、 ACP活性、KR活性、DH活性、以及其等之組合；（b)—多 肽，其包含與序列辨識編號：8有至少80%的同一性之胺基 酸序列，其中該多肽包含KS活性；（c)一多肽，其包含與序 列辨識編號：10有至少80%的同一性之胺基酸序列，其中 該多肽包含MAT活性；（d)—多肽，其包含與序列辨識編號： 14、16、18、20、22，或24的任何一者之有至少80%的同 一性之胺基酸序列，其中該多肽包含ACP活性；（e)—多肽， 其包含與序列辨識編號：12有至少80%的同一性之胺基酸 序列，其中該多肽包含ACP活性；（f)一多肽，其包含與序 列辨識編號：26有至少80%的同一性之胺基酸序列，其中 該多肽包含KR活性；以及(g)—多肽，其包含與序列辨識編 號：28有至少80%的同一性之胺基酸序列，其中該多肽包 含DH活性。於一些具體例中，該胺基酸序列係各別與序列 辨識編號：2、8、10、12、14、16、18、20、22、24、26， 21 201038734 及2 8有至少9 〇 %的同一性或有至少9 5 %的同一性。於一歧呈 - /、 體例中，該等多肽各別包含序列辨識編號：2、8、1〇、12、 14'16、18、20'22、24、26’ 及28之胺基酸序列。 本發明係針對一種經單離的多肽，其係選自於以下所 構成的群組：（a)—多肽，其包含與序列辨識編號：4有至少 80%的同一性之胺基酸序列，其中該多肽包含選自於以下 所構成的群組之PUFA合成酶活性：KS活性、CLF活性、AT 活性、ER活性、以及其等之組合；（b)—多肽，其包含與序 列辨硪編號：30有至少80%的同一性之胺基酸序列，其中 該多肽包含KS活性；（c)一多肽，其包含與序列辨識編號： 32有至少80%的同一性之胺基酸序列，其中該多肽包含clf 活性；（d)—多肽，其包含與序列辨識編號：34有至少80% 的同一性之胺基酸序列，其中該多肽包含Ατ活性；以及(e) 一多肽’其包含與序列辨識編號：36有至少80%的同一性 之胺基酸序列’其中該多肽包含ER活性。於一些具體例中， 該胺基酸序列係各別與序列辨識編號：4、30、32、34，及 36有至少90%的同一性或有至少95%的同一性。於一些具體 例中，該多肽各別包含序列辨識編號_· 4、30、32、34，及 36的胺基酸序列。 本發明係針對一種經單離的多肽，其係選自於以下所 構成的群組：（a)—多肽，其包含與序列辨識編號：6有至少 80%的同一性之胺基酸序列，其中該多肽包含選自於以下 所構成的群組之PUFA合成酶活性：DH活性、ER活性、以 及其等之組合；（b)—多肽，其包含與序列辨識編號：38有 22 201038734 至v 80/。的同-性之胺基酸序列，其中該多肽包含DH活 I·生’⑷夕肽，其包含與序列辨識編號：4〇有至少8〇%的 同-性之胺基酸序列，其中該多&包含训活性 ;以及（d) — 夕肽’其包含與序列辨識編號：42有至少8〇〇/。的同一性之 胺基酸序列，其中該多肽包含ER活性。於一些具體例中， 該胺基酸序列係各別與序列辨識編號：6、38、4〇，及财 至少90〇/〇的同-性或有至少95%的同一性。於一些具體例 〇 中，該等多肽各別包含序列_編號：6、38、40，及42之 胺基酸序列。 本發明係針對-種經單離的多肽，其係選自於以下所 ' 構成的群組：⑷一多肽，其包含與序列辨識編號：69有至 - 外〇%的同一性之胺基酸序列，其中該多肽包含選自於以 下所構成的群組之PUFA合成酶活性：Ks活性、MAT活性、 ACP活性、KR活性、DH活性、以及其等之組合；（b)一多 肽，其包含與序列辨識編號：75有至少8〇%的同一性之胺 〇 基酸序列，其中該多肽包含KS活性；（c)一多肽，其包含與 序列辨識編號：77有至少80%的同一性之胺基酸序列，其 中該多肽包含MAT活性；⑷-多肽’其包含與序列辨識編 號：81、83、85 ' 87 ' 89、91 ' 93、95、97或99的任一者 之有至少80%的同一性之胺基酸序列，其中該多肽包含Acp 活性；（e)—多肽，其包含與序列辨識編號：79有至少8〇% 的同一性之胺基酸序列，其中該多肽包含ACP活性；⑺一 多肽，其包含與序列辨識編號：1〇1有至少80%的同一性之 胺基酸序列，其中該多肽包含KR活性；以及(g)_多狀，其 23 201038734 包含與序列辨識編號：119有至少8〇%的同一性之胺基酸序 列，其中該多肽包含DH活性。於一些具體例中，該胺基酸 序列係各別與序列辨識編號：69、75、77、79、81、83、 85、87、89、91、93、95、97 ' 99、1〇1，以及 119有至少 90 的同一性或有至少95%的同一性。於一些具體例中，該 等多肽各別包含序列辨識編號：69、75、77、79、81、83、 85、87、89、91、93、95、97、99 ' 1(Π，以及 119之胺基 酸序列。 本發明係針對一種經單離的多肽，其係選自於以下所 構成的群組：（a)—多肽，其包含與序列辨識編號：71有至 少80%的同一性之胺基酸序列，其中該多肽包含選自於以 下所構成的群組之PUFA合成酶活性：ks活性、CLF活性、 AT活性、ER活性、以及其等之組合；（b)一多肽，其包含與 序列辨識編號：103有至少80%的同一性之胺基酸序列，其 中該多肽包含KS活性；（c)一多肽，其包含與序列辨識編 號：105有至少80%的同一性之胺基酸序列，其中該多肽包 含CLF活性；（d)—多肽，其包含與序列辨識編號：1〇7有至 少80%的同一性之胺基酸序列，其中該多肽包含AT活性； 以及(e)—多肽，其包含與序列辨識編號：1〇9有至少8〇%的 同一性之胺基酸序列，其中該多肽包含ER活性。於一些具 體例中’該胺基酸序列係與各別與序列辨識編號：B、103、 105、107 ’以及109有至少90°/。的同一性或有至少95%的同 一性。於一些具體例中，該多肽各別包含序列辨識編號： 71、103、105、107，以及109的胺基酸序列。 24 201038734 Ο 本發明係針對一種經單離的多肽，其係選自於以下所 構成的群組：（a)—多肽’其包含與序列辨識編號：73有至 少80%的同一性之胺基酸序列，其中該多肽包含選自於以 下所構成的群組之PUFA合成酶活性：DH活性、ER活性、 以及其等之組合；（b)—多肽，其包含與序列辨識編號：^ 有至少80%的同一性之胺基酸序列，其中該多肽包含〇只活 性；（c) 一多肽’其包含與序列辨識編號：113有至少8〇%的 同一性之胺基酸序列，其中該多肽包含DH活性；以及(句 一多肽，其包含與序列辨識編號：115有至少80%的同—性 之胺基酸序列，其中該多肽包含ER活性。於一些具體例中， 該胺基酸序列係各別與序列辨識編號：73、111、U3，、 及115有至少90%的同一性或有至少95%的同一性。於—此 具體例中，該等多肽各別包含序列辨識編號：73、in、113， 以及115之胺基酸序列。q PUFA synthase activity of the group consisting of: KS activity, CLF activity, AT activity, ER activity, and combinations thereof; (b) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein The polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 30, and wherein the polypeptide comprises KS activity; (c) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein The polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 32, and wherein the polypeptide comprises CLF activity; (d) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein The polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 34, 11 201038734, and wherein the polypeptide comprises AT activity; and (e) a nucleic acid molecule comprising a polynucleotide encoding a polypeptide A sequence wherein the polypeptide comprises an amino acid sequence having at least 80% identity to Sequence ID: 36, and wherein the polypeptide comprises ER activity. In some embodiments, the amino acid sequence is at least 90% identical or at least 95% identical to the sequence identification numbers: 4, 30, 32, 34, and 36, respectively. In some embodiments, the polypeptides each comprise an amino acid sequence of sequence identification numbers: 4, 30, 32, 34, and 36. The present invention is directed to an isolated nucleic acid molecule selected from the group consisting of: 0) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 5, Wherein the polynucleotide sequence encodes a polypeptide comprising PUFA synthase activity selected from the group consisting of DH activity, ER activity, and combinations thereof; (b) a nucleic acid molecule comprising Sequence Identification Number: 37 A polynucleotide sequence having at least 80% identity, wherein the polynucleotide sequence encodes a polypeptide comprising DH activity; (c) a nucleic acid molecule comprising: and a sequence identification number: 39 a polynucleotide sequence of at least 80% identity, wherein the polynucleotide sequence encodes a polypeptide comprising DH activity; and (d) a nucleic acid molecule comprising at least 80% identical to the sequence ID: 41 A polynucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising ER activity. In some embodiments, the polynucleotide sequences are each at least 90% identical to the sequence identification number: 5, 37, 39, and 41 or have at least 95°/. Identity. In some embodiments, the nucleic acid molecules each comprise a polynucleotide sequence of sequence identification numbers: 5, 37, 39, and 41. 12 201038734 The present invention is directed to an isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises a sequence Identification number: 6 an amino acid sequence having at least 80% identity, and wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of: DH activity, ER activity, and combinations thereof; b) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence number: 38, and wherein the polypeptide comprises DH activity; c) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence number: 40, and wherein the polypeptide comprises DH activity; (d) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 42, and wherein the polypeptide Containing ER activity. In some embodiments, the amino acid sequence is at least 90% identical or at least 95% identical to the sequence identification numbers: 6, 38, 40, and 42. In some embodiments, the polypeptides each comprise an amino acid sequence of sequence identification numbers: 6, 38, 40, and 42. The present invention is directed to an isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising at least 80% identical to the sequence identification number: 68 or the sequence identification number: 120. A polynucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising PUFA synthase activity selected from the group consisting of KS activity, MAT activity, ACP activity, KR activity, DH activity, And a combination thereof; (b) - nucleus 13 201038734 an acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 74, wherein the polynucleotide sequence encodes a polypeptide comprising KS activity; (c) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 76, wherein the polynucleotide sequence encodes a polypeptide comprising MAT activity; (d) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to any one of sequence identification numbers: 80, 82, 84, 86, 88, 90, 92, 94, 96 or 98, wherein the multinuclear Glycosidic sequence encoding a polypeptide, its package ACP activity; (e) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 78, wherein the polynucleotide sequence encodes a polypeptide comprising ACP activity; (f) A nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 100, wherein the polynucleotide sequence encodes a polypeptide comprising KR activity; and (g) a nucleic acid molecule A polynucleotide sequence comprising at least 80% identity to sequence identification number: 118, wherein the polynucleotide sequence encodes a polypeptide comprising DH activity. In some embodiments, the polynucleotide sequences are individually and sequence identification numbers: 68, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 118 , and 120 has at least 90 ° /. The identity may have at least 95% identity. In some embodiments, the nucleic acid molecules each comprise a sequence identification number: 68, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 118, and A polynucleotide sequence of 120. The present invention is directed to an isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises a sequence identification number :69 having at least 80% of the amino acid sequence of 201038734 identity, and wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of KS activity, MAT activity, ACP activity, KR activity, DH a combination of activity, and the like; (b) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 75, And wherein the polypeptide comprises KS activity; (c) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 77, And wherein the purine polypeptide comprises MAT activity; (d) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises and sequence identification numbers: 81, 83, 85, 87, 89, 91, 93 Or any one of 95, 97 or 99 having at least 80% identity amino acid sequence, and wherein the polypeptide comprises ACP activity; (e) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, Wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 79, and wherein the polypeptide comprises ACP activity; (f) a nucleic acid molecule, Q comprising a polynucleotide sequence encoding a polypeptide Wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 101, and wherein the polypeptide comprises KR activity; and (g) a nucleic acid molecule comprising a polynucleotide encoding a polypeptide A sequence, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 119, and wherein the polypeptide comprises DH activity. In some specific examples, the amino acid sequence is different from the sequence 四 (4) identification number: 69, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101 And 119 have at least 90% identity or at least 95% identity. In some embodiments, the individual polypeptides 15 201038734 comprise sequence identification numbers: 69, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, and 119 Amino acid sequence. The present invention is directed to an isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising at least 80% identical to the sequence identification number: 70 or sequence identification number: 121 A polynucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising PUFA synthase activity selected from the group consisting of KS activity, chain length factor (CLF) activity, thiol transferase (AT) activity, mercapto-ACP reductase (ER) activity, and combinations thereof; (b) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 102 Wherein the polynucleotide sequence encodes a polypeptide comprising KS activity; (c) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 1〇4, wherein the multinuclear sequence The acid sequence encodes a polymorphism comprising CLF activity; (d) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to the sequence number: 1〇6, wherein the polynucleotide sequence is encoded a polypeptide comprising AT activity, (E) - nucleic acid molecule comprising a sequence identification number: 108 at least 80% identity to a polynucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising an ER activity. In some embodiments, the polynucleotide sequences are each at least 90° per sequence identification number: 7〇, 1〇2, 104, 106, 108, and 121. The identity may have at least 95% identity. In some embodiments, the nucleic acid molecules each comprise a polynucleic acid sequence of sequence identification numbers: 70, 102, 104, 1〇6, 108, and 121. The present invention is directed to an isolated nucleic acid molecule selected from the group consisting of: 16 201038734: (a) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises a sequence Identification number: 71 amino acid sequence having at least 80% identity, and wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of KS activity, CLF activity, AT activity, ER activity', and a combination thereof; (b) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 8% identity with sequence number: 103, and wherein The scorpion polymorphism comprises KS activity; (c) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide wherein the polypeptide comprises at least 8% homozygous amino acid with a sequence number: 105 a sequence, and wherein the polypeptide comprises a CLF-activity' (d)-nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the S-Hole polypeptide comprises at least 80% identity to the sequence ID: 107 It a base acid sequence, and wherein the polypeptide comprises a tau activity; and a nuclear prosthetic knife comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amine having at least 80% identity to sequence identification number: 109 Base acid sequence Ο !·! And wherein the s-like shape contains ER activity. In some embodiments, the amino acid sequence differs from the sequence identification number: 71, 1〇3, 1〇5, 1〇7, and 109 by at least 9% identity or at least %% identical Sex. In some embodiments, the polypeptides each comprise a sequence identity number: 71, 1 〇 3, 1 〇 5, 107, and a 〇 9 amino acid sequence. The present invention is directed to an isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising at least 8% of the sequence identification number: 72 or sequence identification number: 122. A polynucleotide sequence of identity, wherein the polynucleotide sequence encodes a polypeptide comprising PUFA synthase activity selected from the group consisting of: 17 201038734: DH activity, ER activity, and combinations thereof; (b) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 110, wherein the polynucleotide sequence encodes a polypeptide comprising DH activity; (c) a nucleic acid molecule a polynucleotide sequence comprising at least 80% identity to sequence identification number: 112, wherein the polynucleotide sequence encodes a polypeptide comprising DH activity; and (d) a nucleic acid molecule comprising a sequence Identification number: 114 has a polynucleotide sequence of at least 80% identity, wherein the polynucleotide sequence encodes a polypeptide comprising ER activity. In some embodiments, the polynucleotide sequences are each at least 90% identical or at least 95% identical to the sequence identification numbers: 72, 110, 112, 114, and 122. In some embodiments, the nucleic acid molecules each comprise the polynucleotide sequences of sequence identification numbers: 72, 110, 112, 114, and 122. The present invention is directed to an isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises a sequence identification number :73 having at least 80% identity amino acid sequence, and wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of: DH activity, ER activity, and combinations thereof; (b) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 111, and wherein the polypeptide comprises DH activity; (c) A nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 113, and wherein the polypeptide comprises DH activity; and (d) a nucleic acid of 18 201038734 comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 115, and wherein The polypeptide contains ER activity. In some embodiments, the amino acid sequence is at least 90% identical or at least 95% identical to the sequence identification numbers: 73, 111, 113, and 115, respectively. In some embodiments, the polypeptides each comprise an amino acid sequence of sequence identification numbers: 73, 111, 113, and 115. The present invention is directed to an isolated nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide comprising PUFA synthase activity selected from the group consisting of KS activity, MAT activity, ACP activity a combination of KR activity, CLF activity, AT activity, ER activity, DH activity, and the like, wherein the polynucleotide is complementary to any of the polynucleotide sequences described above under stringent conditions Hybridization. The present invention is directed to an isolated nucleic acid molecule comprising a polynucleotide sequence that is fully complementary to any of the polynucleotide sequences set forth above. The present invention is directed to a recombinant nucleic acid molecule comprising: any one of the nucleic acid molecules described above or a combination thereof, and a transcriptional control sequence. In some embodiments, the recombinant nucleic acid molecule is a recombinant vector. The present invention is directed to a host cell which exhibits any of the nucleic acid molecules described above, any of the above-described recombinant nucleic acid molecules, and combinations thereof. In some embodiments, the host cell line is selected from the group consisting of plant cells, microbial cells, and cells. In some embodiments, the microbial cell is a bacterium. In some embodiments, the bacterium is Escherichia coli. In some embodiments, the bacterium is a marine bacterium. In some embodiments, the microbial cell is a Thraustochytrium. In some embodiments, the Thraustochytrium is a Schizochytrium. In some embodiments, the Thraustochytrium is a Thraustochytrium. In some embodiments, the Thraustochytrium is W. Kenyan. (The present invention is directed to a method of producing at least one PUFA comprising: expressing a PUFA synthase gene in a host cell under conditions effective to produce PUFA, Wherein the PUFA synthase gene comprises any one of the above-described isolated nucleic acid molecules, any of the above-described recombinant nucleic acid molecules, or a combination thereof, and at least one of the PUFAs In one aspect of this specific example, the host cell line is selected from the group consisting of: plant cells, isolated animal cells, and microbial cells. In another aspect of this specific example The at least one PUFA comprises docosahexaenoic acid (DHA) or eicosapentaenoic acid (EPA). The present invention is directed to a method of producing a lipid enriched in a combination of DHA, EPA, or a combination thereof, The method comprises: expressing a pUFA synthase gene in the injured host cell under conditions effective for producing a lipid, wherein the PUFA synthase gene comprises any one of the unilaterally isolated nucleic acid molecules described above, the above-described Any of the recombinant nucleic acid molecules, or the like, is combined in a "Herb cell, and a lipid system in which a group of σ is enriched in DHA, or a group thereof. The present invention is directed to a manufacturing A method of recombinant vector comprising inserting any of the above-described isolated nucleic acid molecules into a vector. 20 201038734 The present invention is directed to a method of making a recombinant host cell comprising a recombination The vector vector is introduced into a host cell as described above. In some embodiments, the host cell line is selected from the group consisting of plant cells, isolated animal cells, and microbial cells. Is directed to any of the polynucleotide sequences described above by the encoded isolated polypeptide. The invention is directed to an isolated polypeptide selected from the group consisting of: (a a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 2, wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of: KS activity a combination of MAT activity, ACP activity, KR activity, DH activity, and the like; (b) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 8, wherein the polypeptide comprises KS (c) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 10, wherein the polypeptide comprises MAT activity; (d) - a polypeptide comprising the sequence identification number: Any of amino acid sequences of 14, 16, 18, 20, 22, or 24 having at least 80% identity, wherein the polypeptide comprises ACP activity; (e) - a polypeptide comprising, and a sequence identification number: 12 having at least 80% identity amino acid sequence, wherein the polypeptide comprises ACP activity; (f) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence number: 26, wherein The polypeptide comprises KR activity; and (g) a polypeptide comprising an amino acid sequence having at least 80% identity to Sequence ID: 28, wherein the polypeptide comprises DH activity. In some embodiments, the amino acid sequence differs from the sequence identification number: 2, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 21 201038734 and 2 8 at least 9 〇 The identity of % may have at least 95% identity. In the formula, the polypeptides each comprise a sequence identification number: 2, 8, 1 , 12, 14'16, 18, 20'22, 24, 26' and 28 amino acid sequences . The present invention is directed to a ligated polypeptide selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 4, Wherein the polypeptide comprises a PUFA synthase activity selected from the group consisting of KS activity, CLF activity, AT activity, ER activity, and combinations thereof; (b) - a polypeptide comprising a sequence identification number : 30 having at least 80% identity amino acid sequence, wherein the polypeptide comprises KS activity; (c) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 32, Wherein the polypeptide comprises clf activity; (d) - a polypeptide comprising an amino acid sequence having at least 80% identity to sequence ID: 34, wherein the polypeptide comprises a tau activity; and (e) a polypeptide An amino acid sequence comprising at least 80% identity to sequence identification number: 36 wherein the polypeptide comprises ER activity. In some embodiments, the amino acid sequence is at least 90% identical or at least 95% identical to the sequence identification numbers: 4, 30, 32, 34, and 36, respectively. In some embodiments, the polypeptides each comprise an amino acid sequence of sequence identification numbers _, 4, 30, 32, 34, and 36. The present invention is directed to a ligated polypeptide selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 80% identity to Sequence ID:6, Wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of DH activity, ER activity, and combinations thereof; (b) polypeptide, comprising: and sequence identification number: 38 with 22 201038734 to v 80/. a homo-amino acid sequence, wherein the polypeptide comprises a DH-activated I(4) oxime peptide comprising an amino acid sequence having at least 8% homology to the sequence identification number: 4〇, wherein Multiple & contains training activity; and (d) - oxime peptide 'containing at least 8 〇〇/ with sequence identification number: 42. An amino acid sequence of identity wherein the polypeptide comprises ER activity. In some embodiments, the amino acid sequence is each identical to the sequence identification number: 6, 38, 4, and at least 90 Å/〇 or at least 95% identical. In some embodiments, the polypeptides each comprise an amino acid sequence of sequence number: 6, 38, 40, and 42. The present invention is directed to an isolated polypeptide which is selected from the group consisting of: (4) a polypeptide comprising an amino group having the identity of the sequence identification number: 69 to - outer % An acid sequence, wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of Ks activity, MAT activity, ACP activity, KR activity, DH activity, and combinations thereof; (b) a polypeptide, An amino acid sequence comprising at least 8% identity with sequence identification number: 75, wherein the polypeptide comprises KS activity; (c) a polypeptide comprising at least 80% of the sequence number: 77 An amino acid sequence of the same identity, wherein the polypeptide comprises MAT activity; (4) a polypeptide comprising: any one of sequence identification numbers: 81, 83, 85 '87 '89, 91 '93, 95, 97 or 99 An amino acid sequence having at least 80% identity, wherein the polypeptide comprises Acp activity; (e) a polypeptide comprising an amino acid sequence having at least 8% identity with sequence number: 79, wherein The polypeptide comprises ACP activity; (7) a polypeptide comprising the sequence identification number: 1〇1 to a less than 80% identical amino acid sequence, wherein the polypeptide comprises KR activity; and (g) _ polymorphism, 23 201038734 comprises an amino acid sequence having at least 8 % identity with sequence number: 119 Wherein the polypeptide comprises DH activity. In some embodiments, the amino acid sequence is unique and sequence identification number: 69, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97 '99, 1〇1 And 119 have at least 90 identity or at least 95% identity. In some embodiments, the polypeptides each comprise a sequence identifier number: 69, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99 '1 (Π, and 119 Amino acid sequence. The present invention is directed to an isolated polypeptide selected from the group consisting of: (a) a polypeptide comprising at least 80% identity to sequence identification number: 71. An amino acid sequence, wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of: ks activity, CLF activity, AT activity, ER activity, and combinations thereof; (b) a polypeptide, It comprises an amino acid sequence having at least 80% identity to sequence identification number: 103, wherein the polypeptide comprises KS activity; (c) a polypeptide comprising at least 80% identity to sequence identification number: 105 An amino acid sequence, wherein the polypeptide comprises CLF activity; (d) a polypeptide comprising an amino acid sequence having at least 80% identity to the sequence number: 1〇7, wherein the polypeptide comprises AT activity; (e) a polypeptide comprising at least 8% identical to the sequence identification number: 1〇9 An amino acid sequence, wherein the polypeptide comprises ER activity. In some embodiments, the amino acid sequence has at least 90° with each of the sequence identification numbers: B, 103, 105, 107' and 109. Identity may have at least 95% identity. In some embodiments, the polypeptides each comprise an amino acid sequence of sequence identification numbers: 71, 103, 105, 107, and 109. 24 201038734 Ο The present invention is directed to a An isolated polypeptide selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 73, wherein the polypeptide comprises PUFA synthase activity from the group consisting of: DH activity, ER activity, and combinations thereof; (b) a polypeptide comprising an amino group having at least 80% identity to the sequence identification number: An acid sequence, wherein the polypeptide comprises 〇-only activity; (c) a polypeptide comprising an amino acid sequence having at least 8% identity with sequence identification number: 113, wherein the polypeptide comprises DH activity; a polypeptide comprising a sequence identification number 115 has at least 80% homologous amino acid sequence, wherein the polypeptide comprises ER activity. In some embodiments, the amino acid sequence is unique and sequence identification number: 73, 111, U3, and 115 There is at least 90% identity or at least 95% identity. In this particular example, the polypeptides each comprise a sequence identity number: 73, in, 113, and 115 amino acid sequence.

於一些具體例中，本發明之經單離的多肽的任— 以是一融合多肽。 者可 本發明係針對一組成物，其包含以上說明的多肽之任 一者以及生物上可接受的載體。 本發明係針對一種於具有p UFA合成酶活性的有機體 内增加DHA、EPA，或其等之一組合的生產的方法，其包 含：於有效產生DHA、EPA或其等之一組合的條件下於— 有機體中表現以上說明的該等經單離的核酸分子的任— 者、以上說明的該等重組型核酸分子的任一者，或其等之 組合，其中該PUFA合成酶活性取代該有機體中之不活化咬 25 201038734 被刪除的雜，引人—新的活性，或提高畴的活性，以 及其中增加該有機體中的DHA、EPA，或其等之一組合之 生產。In some embodiments, any of the isolated polypeptides of the invention is a fusion polypeptide. The present invention is directed to a composition comprising any of the polypeptides described above and a biologically acceptable carrier. The present invention is directed to a method for increasing production of a combination of DHA, EPA, or the like in an organism having p UFA synthetase activity, comprising: under conditions effective to produce a combination of DHA, EPA, or the like Or the combination of any of the above-described isolated nucleic acid molecules described above, any of the above-described recombinant nucleic acid molecules, or a combination thereof, wherein the PUFA synthase activity is substituted for the organism Non-activated bite 25 201038734 Deletion of impurities, introduction of new activity, or enhancement of domain activity, and production of a combination of DHA, EPA, or the like in the organism.

本發明係針對一種自宿主細胞單離脂質的方法，其包 3 (a)於有效產生脂質的條件下於該宿主細胞中表現puFA 合成酶基因’其中該PUFA合成酶基因包含以上說明的該等 、.二單離的核酸分子的任一者、以上說明的該等重組型核酸 y刀子的任—者，或其等之組合於該宿主細胞中，以及(b)自 α亥佰主細胞單離脂質。於一些具體例中，該宿主細胞係選 自於以下所構成的群組：植物細胞、經單離的動物細胞， 以及微生物細胞。於一些具體例中，該等脂質包含£)11八、 ΕΡΑ ’或其等之—組合。 圖式簡單說明 第1圖顯示出本發明的裂殖壺菌種ATCC ΡΤΑ_9695 PUFA合成酶之基因架構； 第2圖顯示出本發明的破囊壺菌種ATCC PTA-10212 PUFA合成酶之基因架構； 第3圖顯示出本發明的裂殖壺菌種ATcc PTA-9695和 破囊壺菌種ATCC PTA-10212 PUFA合成酶以及來自裂殖壺 菌種ATCC 20888、破囊壺菌種ATCC 20892、金黄色破囊壺 菌，和SAM2179之合成酶之領域架構； 第4圖顯示出本發明裂殖壺菌種ATCC PTA-9695 Pfalp 胺基酸序列（序列辨識編號：2)和破囊壺菌種ATCC PTA-10212 Pfalp胺基酸序列（序列辨識編號：69)的排列， 26 201038734 其等具有該等之來自裂殖壺菌種ATCC 20888 (序列辨識編 號：54)和破囊壺菌種ATCC 20892 (序列辨識編號：56)的 OrfA序列以及來自金黄色破囊壺菌（序列辨識編號：55)的 OrfA序列； 第5圖顯示出排列本發明的裂殖壺菌種ATCC PTA-9695 Pfa2p胺基酸序列(序列辨識編號：4)和破囊壺菌 種ATCC PTA-10212 Pfa2p胺基酸序列（序列辨識編號：71) 的排列，其等具有該等之來自裂殖壺菌種ATCC 20888 (序 列辨識編號：57)和破囊壺菌種ATCC 20892 (序列辨識編 號：58)的Orffi序列以及來自金黄色破囊壺菌（序列辨識編 號：59)的OrfB序列； 第6圖顯示本發明的裂殖壺菌種ATCC PTA-9695 Pfa3p 胺基酸序列（序列辨識編號：6)以及破囊壺菌種ATCC PTA-10212 Pfa3p胺基酸序列（序列辨識編號：73)的排列， 其等具有來自裂殖壺菌種ATCC 20888 (序列辨識編號：61) 以及破囊壺菌種ATCC 20892 (序列辨識編號：60)的OrfC序 列； 第7圖顯示裂殖壺菌種ATCC PTA-9695 多核苷酸 序列(序列辨識編號：1); 第8圖顯示裂殖壺菌種ATCC PTA-9695 Pfalp胺基酸序 列(序列辨識編號：2); 第9圖顯示裂殖壺菌種ATCC PTA-9695 多核苷酸 序列(序列辨識編號：3); 第10圖顯示裂殖壺菌種ATCC PTA-9695 Pfa2p胺基酸 27 201038734 序列（序列辨識編號：4); 第11圖顯示裂殖壺菌種ATCC PTA-9695 PM3多核苷 酸序列（序列辨識編號：5); 第12圖顯示裂殖壺菌種ATCC PTA-9695 Pfa3p胺基酸 序列(序列辨識編號：6); 第13圖顯示破囊壺菌種ATCC PTA-10212尸柯7多核苷 酸序列（序列辨識編號：68); 第14圖顯示破囊壺菌種ATCC ΡΤΑ-1〇212户柯/多核苷 酸序列（序列辨識編號：120)，其已經為了於裂殖壺菌内表 現而被密碼子最佳化； 第I5圖顯示破囊壺菌種ATCC PTA-10212 Pfalp胺基酸 序列（序列辨識編號：69); 第16圖顯示破囊壺菌種ATCC PTA-10212户抑2多核苷 酸序列（序列辨識編號：70); 第17圖顯示破囊壺菌種ATCC PTA-10212 多核苷 酸序列（序列辨識編號：121)，其已經為了於裂殖壺菌内表 現而被密碼子最佳化； 第18圖顯示破囊壺菌種ATCC PTA-10212 Pfa2p胺基酸 序列（序列辨識編號：71); 第19圖顯示第13圖顯示破囊壺菌種ATCC PTA-10212 多核苷酸序列(序列辨識編號：72); 第20圖顯示破囊壺菌種ATCC ΡΤΑ-10212尸Ε43多核苷 酸序列（序列辨識編號：122)，其已經為了於裂殖壺菌内表 現而被密碼子最佳化； 28 201038734 第21圖顯示破囊壺菌種ATCC PTA-10212 Pfa3p胺基酸 序列（序列辨識編號：73); 第22圖顯示裂殖壺菌的密碼子使用表。 【實施冷式】 較佳實施例之詳細説明 本發明係針對涉及PUFAs的生產之多不飽和脂肪酸 (PUFA)合成酶的經單離的核酸分子及多肽，包括富含二十 二碳六烯酸(DHA)、二十碳五烯酸(EPA)，或其等之一組合 的PUFAs。本發明係針對包含該等核酸分子的載體和宿主 細胞、由該等核酸分子編碼的多肽、包含該等核酸分子或 多肽的組成物，以及製造其等之方法和其等之用途。 PUFA合成酶 當使用於本文中，術語，，PUFA合成酶"係提及涉及生產 多不飽和脂肪酸的一酵素。參見，例如，Metz等人，Science 293:290-293 (2001)。 本發明在某種程度上係針對3種PUFA合成酶次單元， 稱為Pfalp(序列辨識編號：2或序列辨識編號：69)、Pfa2p (序 列辨識編號：4或序列辨識編號：71)，和Pfa3p (序列辨識 編號：6或序列辨識編號：73)，以及編碼該等次單元之基 因，稱為尸柯7 (序列辨識編號：1、序列辨識編號：68，或 序列辨識編號：120)、尸抱2 (序列辨識編號：3、序列辨識 編號：70,或序列辨識編號：121)，和P柯3 (序列辨識編號： 5、序列辨識編號：72，或序列辨識編號：122)。參見，第 1-3圖與第7-21圖。於其他破囊壺菌内的PUFA合成酶亦已經 29 201038734 各別地稱為ORF 1、ORF 2，和ORF 3，或是各別地稱為 OrfA、orfB，和orfC。參見，例如：美國專利案第7,247,461 號和第7,256,022號中的裂殖壺菌種(ATCC 20888)以及破囊 壺菌種(ATCC 20892)，參照orfA、orfB ’和orfC基因及對應 的OrfA、orfB，和orfC蛋白質，以及美國專利案第7,368,552 號中的金黄色破囊壺菌（ATCC 34304)，參照〇RF A、ORF B ’和ORF C基因與蛋白質。亦可參見，WO/2005/097982 中的菌株SAM2179，參照ORF 1、ORF 2，和ORF 3基因與 蛋白質。 核酸分子 本發明係針對經單離的核酸分子，其包含自一經單離 的微生物所衍生的PUFA合成酶基因與領域的多核苷酸序 列，其係2009年3月19曰提申之共審查美國申請案第 12/407,687號的主題，以其之整體併入本文中作為參考資 料。s玄微生物係依據布達佩斯條約於2〇〇9年丨月7日被寄存 於美國類型培養物收集中心，專利寄存’ 1〇8〇1 UnWersity Boulevard，Manassas，VA 2〇11〇_22〇9，以及授予atcc寄存 編號 PTA-9695The present invention is directed to a method for isolating a lipid from a host cell, the kit 3 (a) expressing a puFA synthase gene in the host cell under conditions effective to produce a lipid, wherein the PUFA synthase gene comprises the above-described Any one of the two isolated nucleic acid molecules, any of the above-described recombinant nucleic acid y knives, or the like, or the combination thereof, and (b) from the α 佰 佰 细胞From lipids. In some embodiments, the host cell line is selected from the group consisting of plant cells, isolated animal cells, and microbial cells. In some embodiments, the lipids comprise a combination of £11, ’' or the like. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows the genetic architecture of the Schizochytrium sp. ATCC ΡΤΑ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Figure 3 shows the Schizochytrium sp. ATcc PTA-9695 and the Thraustochytrium sp. ATCC PTA-10212 PUFA synthase and the Schizochytrium sp. ATCC 20888, the Thraustochytrium sp. ATCC 20892, golden yellow. Thraustochytrium, and the domain architecture of the synthetase of SAM2179; Figure 4 shows the Schizochytrium sp. ATCC PTA-9695 Pfalp amino acid sequence (SEQ ID NO: 2) and the Thraustochytrium ATCC PTA -10212 arrangement of the Pfalp amino acid sequence (SEQ ID NO: 69), 26 201038734 which has the same from Schizochytrium sp. ATCC 20888 (SEQ ID NO: 54) and Thraustochytrium sp. ATCC 20892 (sequence Identification number: 56) OrfA sequence and OrfA sequence from Thraustochytrium aureus (SEQ ID NO: 55); Figure 5 shows arrangement of the Schizochytrium sp. ATCC PTA-9695 Pfa2p amino acid sequence of the present invention (Sequence ID: 4) and Thraustochytrium ATC The arrangement of the C PTA-10212 Pfa2p amino acid sequence (SEQ ID NO: 71), which has the same from the Schizochytrium sp. ATCC 20888 (SEQ ID NO: 57) and the Thraustochytrium sp. ATCC 20892 (sequence) Identification number: 58) Orffi sequence and OrfB sequence from Thraustochytrium aureus (SEQ ID NO: 59); Figure 6 shows Schizochytrium sp. ATCC PTA-9695 Pfa3p amino acid sequence of the present invention (sequence Identification number: 6) and the arrangement of the Thraustochytrium sp. ATCC PTA-10212 Pfa3p amino acid sequence (SEQ ID NO: 73), which has the origin from the Schizochytrium sp. ATCC 20888 (SEQ ID NO: 61) and OrfC sequence of the ampulla of the genus ATCC 20892 (SEQ ID NO: 60); Figure 7 shows the polynucleotide sequence of the Schizochytrium strain ATCC PTA-9695 (SEQ ID NO: 1); Figure 8 shows Schizochytrium ATCC PTA-9695 Pfalp amino acid sequence (SEQ ID NO: 2); Figure 9 shows Schizochytrium sp. ATCC PTA-9695 polynucleotide sequence (SEQ ID NO: 3); Figure 10 shows fission pot Strains ATCC PTA-9695 Pfa2p Amino Acid 27 201038734 Sequence (Sequence Identification Figure 4 shows the ATCC PTA-9695 PM3 polynucleotide sequence of Schizochytrium sp. (SEQ ID NO: 5); Figure 12 shows the Schizochytrium sp. ATCC PTA-9695 Pfa3p amino acid sequence (sequence Identification number: 6); Figure 13 shows the cysticidopsis species ATCC PTA-10212 nectar 7 polynucleotide sequence (SEQ ID NO: 68); Figure 14 shows the Thraustochytrium strain ATCC ΡΤΑ-1〇212 Co/polynucleotide sequence (SEQ ID NO: 120), which has been codon-optimized for expression in Schizochytrium; Figure I5 shows Thraustochytrium ATCC PTA-10212 Pfalp amino acid sequence (SEQ ID NO: 69); Figure 16 shows the ATCC PTA-10212-inhibited 2 polynucleotide sequence of the Thraustochytrium sp. (Sequence ID: 70); Figure 17 shows the Thraustochytrium ATCC PTA-10212 multinuclei Glycosidic acid sequence (SEQ ID NO: 121), which has been codon optimized for expression in Schizochytrium; Figure 18 shows the sequence of Thraustochytrium ATCC PTA-10212 Pfa2p amino acid (sequence identification) No.: 71); Figure 19 shows that Figure 13 shows the polynucleotide sequence of the Thraustochytrium sp. ATCC PTA-10212 (sequence identification) No.: 72); Figure 20 shows the 43-polynucleotide sequence of the Thraustochytrium sp. ATCC ΡΤΑ-10212 corpse (SEQ ID NO: 122), which has been codon-optimized for expression in Schizochytrium; 28 201038734 Figure 21 shows the Thraustochytrium strain ATCC PTA-10212 Pfa3p amino acid sequence (SEQ ID NO: 73); Figure 22 shows the codon usage table for Schizochytrium. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention is directed to isolated nucleic acid molecules and polypeptides comprising polyunsaturated fatty acid (PUFA) synthetase produced by PUFAs, including docosahexaenoic acid-rich PUFAs in combination with (DHA), eicosapentaenoic acid (EPA), or a combination thereof. The present invention is directed to vectors and host cells comprising such nucleic acid molecules, polypeptides encoded by such nucleic acid molecules, compositions comprising such nucleic acid molecules or polypeptides, methods of making the same, and the like. PUFA Synthetase As used herein, the term PUFA synthase refers to an enzyme involved in the production of polyunsaturated fatty acids. See, for example, Metz et al., Science 293: 290-293 (2001). The present invention is directed to some extent to three PUFA synthetase subunits, referred to as Pfalp (SEQ ID NO: 2 or Sequence ID: 69), Pfa2p (SEQ ID NO: 4 or Sequence ID: 71), and Pfa3p (SEQ ID NO: 6 or Sequence ID: 73), and the gene encoding the subunit, called necco 7 (sequence identification number: 1, sequence identification number: 68, or sequence identification number: 120), Corpse 2 (sequence identification number: 3, sequence identification number: 70, or sequence identification number: 121), and P co3 (sequence identification number: 5, sequence identification number: 72, or sequence identification number: 122). See, Figures 1-3 and Figures 7-21. PUFA synthase in other Thraustochytrium has also been referred to as ORF 1, ORF 2, and ORF 3, respectively, or as OrfA, orfB, and orfC, respectively. See, for example, Schizochytrium species (ATCC 20888) and Thraustochytrium species (ATCC 20892) in U.S. Patent Nos. 7,247,461 and 7,256,022, with reference to orfA, orfB' and orfC genes and corresponding OrfA, orfB And orfC proteins, as well as the Thraustochytrium aureus (ATCC 34304) in U.S. Patent No. 7,368,552, with reference to 〇RF A, ORF B ' and ORF C genes and proteins. See also, strain SAM2179 in WO/2005/097982, with reference to ORF 1, ORF 2, and ORF 3 genes and proteins. Nucleic Acid Molecules The present invention is directed to an isolated nucleic acid molecule comprising a PUFA synthase gene and a domain polynucleotide sequence derived from an isolated microorganism, which is reviewed in the United States on March 19, 2009. The subject matter of the application Serial No. 12/407, 687, which is incorporated herein by reference in its entirety. The smectic microbes were deposited with the American Type Culture Collection Center on the 7th of July, 2009, under the Budapest Treaty, and the patent registration '1〇8〇1 UnWersity Boulevard, Manassas, VA 2〇11〇_22〇9, And grant atcc registration number PTA-9695

以及也被稱為裂殖壺菌種ATCC PTA-9695。當表現時，此等基因產生獨特的脂肪酸形態， 係特徵部分在於高位準的亞米加_3脂肪酸，特別是高位準 的 DHA。 本發明係針對經單離的核酸分子，其包含自一經單離 的微生物所衍生的PUFA合成酶基因與領域的多核發酸序 列’其係2010年1月19日提申之共審查美國申請案第 30 201038734 61/296,456號的主題’以其之整體併人本文中作為參考資 料。δ亥微生物係依據布達佩斯條約於2〇〇9年〗月14 曰被寄存 於美國類型培養物收集中心，專利寄存，1〇8〇1 UniverskyAlso known as the Schizochytrium strain ATCC PTA-9695. When expressed, these genes produce a unique fatty acid morphology characterized by high levels of sub-male-3 fatty acids, particularly high-level DHA. The present invention is directed to an isolated nucleic acid molecule comprising a PUFA synthase gene derived from an isolated microorganism and a multi-nucleic acid sequence of the field 'a review of the US application filed on January 19, 2010 The subject matter of the 30th 201038734 61/296,456 is hereby incorporated by reference in its entirety. The δHai Microorganisms were deposited under the Budapest Treaty at 2〇〇9, month 14 at the American Type Culture Collection Center, Patent Depository, 1〇8〇1 Universky

Boulevard，Manassas, VA 2011〇_22〇9，以及授予ATCC寄存 編號PTA-10212，以及也被稱為破囊壺菌種ATCC PTA-10212。當表現時，此等基因產生獨特的脂肪酸形態， 係特徵部分在於高位準的亞米加_3脂肪酸，特別是高位準 的DHA、EPA，或其等之一組合。 當使用於本文中，”多核苷酸"能包含一慣見的磷酸二 酯鍵或慣見的鍵(例如，一醯胺鍵，如，於肽核酸(pNA)中 發現的）。一個多核苷酸可含有全長cDNA序列的核苷酸序 列’包括未轉譯5'和3'序列、編碼序列，以及該核酸序列之 片段、抗原決定位、領域，和變異體。該多核苷酸可以是 由任何的聚核醣核苷酸或是聚去氧核醣核苷酸組成的，其 可以是未經修飾的RNA或DNA或是經修飾的RNA或 DNA。舉例而言，多核苷酸可以是由以下組成的：單股或 雙股的DNA、單股或雙股區域的混合物之DNA、單股或雙 股的RNA，以及單股或雙股區域的混合物之RNA，雜交分 子，其包含可以是單股的DNA和RNA或更典型地雙股的 DNA和RNA，或是單股或雙股區域的混合物。此夕卜，該等 多核苷酸可以是由三股區域組成的，其包含RNA或DNA或 是RNA與DNA二者。多核苷酸可含有核糖核苷(腺苷 '鳥苦、 尿苷，或是胞苷；"RNA分子”)或是去氧核糖核苷（去氧腺苦、 去氧鳥苷、去氧胸腺核苷，或是去氧胞苷；"DNA分子，，）， 31 201038734 或是其等之任何的磷酯類似物，例如··硫代磷酸酯和硫酯。 多核普酸亦可含有〜或更多個經修飾的驗基或是為了安定 性或其他原因而修飾的DNA或RNA主鏈。，，經修飾的，，鹼基 包括’舉例而言’三苯曱基化(tritylated)的驗基和不常見的 鹼基，例如：肌苷。DNA和RNA可以做各種各樣的修飾； 因而，”多核苷酸"包括化學上、酵素上，或是代謝上修飾 的形式。術語核酸分子僅提及分子的一級和二級結構，以 及不限於任何的特定的第三形式。因而，此術語包括發現 的雙股DNA，尤其，線形或是環形的DNA分子(例如：限制 性片段）、質體，以及染色體。於討論的特定的雙股DNA分 子結構中’序列可以於本文中依據僅於沿著DNA的非轉錄 股之5,至3’方向的序列規定的正常的規範來說明（亦即，具 有與mRNA有同源性的序列之該股）。 術語”經單離的"核酸分子係提及一核酸分子、DNa^ RNA ,其已經自其之天然的環境被移除的。經單離的核酸 分子之進一步的實例包括核酸分子，其包含被維持於異源 性宿主細胞中的重組型多核苷酸或是溶液中之經純化的 (部分地或是大體上）多核苷酸。經單離的RNA分子包括本發 明的多核苷酸之活體内或是活體外RNA轉錄品。如本發明 之經單離的核酸分子進一步包括合成地生產的此等分子。 此外，一核酸分子或多核苷酸能包括一調節要素，例如. 一啟動子、核糖體結合位址，或是一轉錄終止子。 一"基因"係提及核苷酸的總成，其編碼一多肽以及 包括cDNA和基因體DNA核酸。，，基因”也提及—核酸片段， 32 201038734 ,、表現特疋的蛋白質，包括介於個別的編碼區段(外顯子) 之間的介人序列（内含子），以及在編碼序列之前的(5,非編 碼序列）以及在編碼序列之後的(3,非編碼序列)調控序列。" 天然的基因係提及―基因，當在自然界發現且帶有其自身 的調控序列。 本發明係針對經單離的核酸分子，其包含與以下有至 V 80/〇同性的多核苷酸序列：裂殖壺菌種atcC PTA-9695 (序列辨哉編號.1)、裂殖壺菌種ATcc pta-9695 (序列辨識編號：3)、裂殖壺菌種ATCC PTA-9695 (序 列辨識編號：5)、破囊壺菌種ATCCPTA-l〇212Pi^7(序列 辨識編號：68或序列辨識編號：12〇)、破囊壺菌種ATCC PTA-10212 (序列辨識編號：70或序列辨識編號： 121)、破囊壺菌種ATCCPTA_10212PFw(序列辨識編號： 72或序列辨識編號：122)的多核苷酸序列，以及其等之組 合’其中該等多核苷酸編碼包含一或更多PUFA合成酶活性 的多肽。 該等PUFA合成酶活性係與各合成酶多肽的一或更多 個領域有關聯，其中該等領域能藉由其等之守恆性結構或 功能性要素(根據其等與已知的要素之同源性）予以鑑定，以 及也可以根據其等之特定的生化活性予以鑑定。參見，例 如，美國專利案第7,217,856號，以其之整體併入本文中作 為參考資料。PUFA合成酶領域之實例包括：Pfalp内的β-酮基醯基-ACP合成酶(KS)領域、丙二醯基-CoA : ACP醯基 轉移酶(MAT)領域、醯基載體蛋白質(ACP)領域、酮還原酶 33 201038734 (KR)領域，和β-經基Si基-ACP脫水酶(DH)領域；pfa2p内的 KS領域、鏈長度因子(CLF)領域、醯基轉移酶(AT)領域，以 及和且稀醯基-ACP還原酶(ER)領域in ;以及pfa3p内的DH 領域與ER領域。 具有β-酮基醯基-ACP合成酶(KS)生物活性（功能）之一 多肽或一多肽的領域先前已經顯示出能夠進行該脂肪酸的 延長反應循環之起始步驟。術語”β-酮基醯基_ACP合成酶" 已經與術語”3-酮基醯基-ACP合成酶'，、"β-酮基醯基-ACP合 成酶"，以及"酮基醯基，ACP合成酶，，互相交換使用。於其他 系統中’已經顯示出用於延長之醢基係藉由一硫酯鍵連結 至KS的活性位置之一個半胱胺酸殘基，以及醯基_ks係與 丙二醯基-ACP進行縮合作用以形成_酮基醯基—Acp、c〇2 與未結合（"自由”)KS。於此等系統中，已經顯示出KS擁有 比反應循環中之其他多肽更大之受質特異性。多肽（或多肽 的領域）能藉由與已知之KS序列之同源性而容易地被鑑定 出為屬於該KS家族。 具有丙二醯基-CoA : ACP醯基轉移酶(MAT)活性之一 多狀或一多肽的領域先前已經顯示出能夠自丙二醯基-CoA 上將丙二醯基部分轉移至ACP。術語，，丙二醯基-CoA : ACP 醯基轉移酶”已經與”丙二醯基醯基轉移酶”互相交換使用。 除了該活性位置要素(GxSxG)之外，已經顯示出MATs擁有 一延伸的要素(於關鍵位置的R與Q胺基酸）。多肽（或多肽的 領域）能藉由其等與已知之mat序列之同源性以及其等之 延伸的要素結構而容易地被鑑定出為屬於該mat家族。 34 201038734 具有醯基載體蛋白質（ACP)活性之一多肽或一多肽的 領域先前已經顯示出能夠作用為成長脂肪醯基鏈之一載 體，其係經由一硫酯鍵聯至共價性結合之輔助因子。ACPs 典型地為大約80至大約1〇〇個胺基酸長以及已經顯示出係 藉由轉移CoA上之攝酸泛疏醇部分（ph〇Sph〇pantetheinyl moiety)至ACP上之高度守恆性絲胺酸殘基而自未活化之不 完全(apo)-形式轉換為功能性完全(h〇i〇)_形式。也已經顯示 出醯基係藉由在磷酸泛硫醇部分的游離端之一硫酯鍵聯而 連結至ACPs。活性位置要素(LGIDS*)變化之存在也已經認 定為ACP之一特徵。該活性位置絲胺酸(S*)的官能性已經於 一細菌的PUFA合成酶内展現出（Jiang等人，j. Am. Chem. Soc. 130:6:336-7 (2〇〇8))。多肽（或多肽的領域)能藉由用放射 性的泛硫醇予以標記及藉由與已知之ACPs之序列同源性 而容易地被鑑定出為屬於該ACP家族。 具有脫水酶（dehydrase)或脫水酶（dehydratase)(DH)活 性之一多肽或一多肽的領域先前已經顯示出能夠催化一脫 水反應。提及DH活性典型地係提及FabA-類似β-羥基醯基 -ACP脫水酶生物活性。FabA-類似β-羥基醯基-ACP脫水酶 生物活性會自β-酮基醯基-ACP移除Η0Η，且一開始會在碳 鏈上製造出一反式雙鍵。術語"FabA-類似β-羥基醯基-ACP 脫水酶"已經與術語"FabA-類似β-羥基醯基-ACP脫水酶·’、 πβ-羥基醯基-ACP脫水酶"及”脫水酶"互相交換使用。PUFA 合成酶系統之DH領域先前已經展現出顯示與FAS系統（而 非其他PKS系統之DH領域）關聯的之細菌DH酵素具有同源 35 201038734 性。參見，例如，美國專利案第7,217,856號，以其之整體 併入本文中作為參考資料。次族群之細菌DH，該FabA-類 似DH，擁有順式-反式異構酶活性（Heath等人，】.6丨〇1· Chem·，271, 27795 (1996))。根據與FabA-類似DH蛋白質之 同源性，該PUFA合成酶系統DH領域的一者或全體可負責 PUFA合成酶產物内之順式雙鍵之插入。一多肽或領域亦能 具有非-FabA-類似DH活性、或是非-FabA-類似β-羥基醯基 -ACP脫水酶(DH)活性。更明確地，PUFA合成酶DH領域中 先前已經鑑定出大約13個胺基酸長的守恆性活性位置要素 之：LxxHxxxGxxxxP (該要素中的L位置也可以是I)。參見， 例如：美國專利案第7,217,856號，以及DonadioS, KatzL.， Gene 111(1):51-60 (1992)，其之各個係以其之整體併入本文 中作為參考資料。此守恆性要素係於全部已知的PUFA合成 酶序列的相似區域中找到以及可以是非-FabA類似脫水作 用的原由。 具有/?-酮基醯基-ACP還原酶(KR)活性之一多肽或一 多肽的領域先前已經顯示出能夠催化3-酮基醯基形式之 ACP之吡啶-核苷酸-依賴型還原作用。術語”万-酮基醯基 -ACP還原酶"已經與術語"酮還原酶"、"3-酮基醯基-ACP還 原酶''，及”酮基-醯基ACP還原酶”互相交換使用。已經被決 定其他系統中的KR功能涉及脂肪酸重新生合成延長循環 之第一還原步驟。多肽（或多肽的領域）能藉由與已知之 PUFA合成酶KRs之序列同源性而容易地被鑑定出為屬於該 KR家族。 36 201038734 具有鏈長度因子（CLF)活性之一多肽或一多肽的領域 先前已經被定義成具有下列的活性或特性的一或更多者： (1) 其能決定延長循環的數目且因此決定終產物的鏈長度， (2) 其具有與KS的同源性’但是缺少KS活性位置半胱胺酸， (3) 其能與KS異體二聚化的，（4)其能提供待延長的起始驗 基，或是(5)其能將丙二酸酯（為丙二醯基-ACP)去叛基，因 而形成可轉移至KS活性位置之醋酸鹽基團以及可作為進行 〇 起始延長(縮合)反應之‘啟動(priming)，分子。在所有最近辨 識出之PUFA合成酶系統中皆發現到CLF領域，以及在每— 情況下皆發現為多領域蛋白質之一部分。多肽（或多肽的領 - 域）能藉由與已知之PUFA合成酶CLFs之序列同源性而容易 . 地被鑑定出為屬於該CLF家族。 具有醯基轉移酶(AT)活性之一多肽或一多肽的領域先 前已經被定義成具有下列的活性或特性之一或更多者：（1) 其能轉移該將脂肪醯基自ACP領域轉移至水（亦即，硫酯 〇 酶），將該脂肪醯基釋放為一游離脂肪酸，（2)其能將一脂肪 醯基轉移至一接受體，例如：CoA，（3)其能轉移各種各樣 的ACP領域之中的醯基，或是(4)其能將脂肪醯基轉移至一 親脂性的接受體分子（例如，至溶血磷脂酸(lys〇ph〇sphadic acid))。多肽（或多肽的領域）能藉由與已知之pUFA合成酶 ATs之序列同源性而容易地被鑑定出為屬於該AT家族。 具有稀醯基-ACP還原酶(ER)生物活性之一多肽或一多 肽的領域先前已經顯示出能夠還原脂肪醯基—ACP上之反式 雙鍵（由DH活性引入）’導致關聯的碳之飽和^ puFA合成酶 37 201038734 系統内的^尺領域先前已經顯示出與ER酵素家族具有同源 性(Heath等人，Nature 406: 145-146 (2000)，以其之整體併 入本文中作為參考資料），以及一ER同源物已經顯示出作用 為活體外之烯酿基-ACP還原酶(Bumpus等人，了.八„1.〇1抓·Boulevard, Manassas, VA 2011〇_22〇9, and the ATCC deposit number PTA-10212, and also known as the Thraustochytrium ATCC PTA-10212. When expressed, these genes produce unique fatty acid forms that are characterized in part by high levels of sub-male-3 fatty acids, particularly high levels of DHA, EPA, or a combination thereof. As used herein, a "polynucleotide" can comprise a conventional phosphodiester bond or a conventional bond (eg, a guanamine bond, such as found in peptide nucleic acids (pNA)). A nucleotide sequence that can contain a full-length cDNA sequence includes both untranslated 5' and 3' sequences, coding sequences, and fragments, epitopes, domains, and variants of the nucleic acid sequences. Any of the polyribonucleotides or polydeoxyribonucleotides, which may be unmodified RNA or DNA or modified RNA or DNA. For example, the polynucleotide may be composed of the following Single or double stranded DNA, a mixture of single or double stranded DNA, single or double stranded RNA, and a mixture of single or double stranded regions of RNA, hybrid molecules, which may be single stranded DNA and RNA or more typically double-stranded DNA and RNA, or a mixture of single or double-stranded regions. In addition, the polynucleotides may be composed of three regions, which comprise RNA or DNA or RNA and DNA. Polynucleotides may contain ribonucleosides (adenosine 'birds , uridine, or cytidine; "RNA molecule") or deoxyribonucleoside (deoxyadenosine, deoxyguanosine, deoxythymidine, or deoxycytidine; "DNA molecules ,,), 31 201038734 Or any of the phospholipid analogs, such as · · phosphorothioates and thioesters. Polynucleotides can also contain ~ or more modified assays or DNA or RNA backbones modified for stability or other reasons. , modified, base includes, for example, a tritylated test group and an unusual base such as inosine. DNA and RNA can be modified in a variety of ways; thus, "polynucleotide" includes chemically, enzymatically, or metabolically modified forms. The term nucleic acid molecule refers only to the primary and secondary structure of the molecule, and Limited to any particular third form. Thus, the term includes the discovered double-stranded DNA, in particular, linear or circular DNA molecules (eg, restriction fragments), plastids, and chromosomes. The 'sequences in the DNA molecular structure can be described herein in terms of normal specifications specified only for sequences along the 5, 3' direction of the non-transcribed strands of DNA (ie, sequences having homology to mRNA) The term "isolated" nucleic acid molecules refers to a nucleic acid molecule, DNa^ RNA, which has been removed from its natural environment. Further examples of isolated nucleic acid molecules include nucleic acid molecules comprising a recombinant polynucleotide maintained in a heterologous host cell or a purified (partially or substantially) polynucleotide in solution . The isolated RNA molecules include in vivo or in vitro RNA transcripts of the polynucleotides of the invention. The isolated nucleic acid molecules of the invention further comprise such molecules produced synthetically. In addition, a nucleic acid molecule or polynucleotide can include a regulatory element, such as a promoter, a ribosome binding site, or a transcription terminator. A "gene" refers to a nucleotide assembly that encodes a polypeptide as well as nucleic acids including cDNA and genomic DNA. , the gene" also refers to - a nucleic acid fragment, 32 201038734, a protein that exhibits amnesia, including a human sequence (intron) between individual coding segments (exons), and in the coding sequence The previous (5, non-coding sequence) and the (3, non-coding sequence) regulatory sequences following the coding sequence. " The natural gene line refers to the gene, when found in nature and carries its own regulatory sequences. The invention is directed to an isolated nucleic acid molecule comprising a polynucleotide sequence having the same V 80 / 〇 homology as follows: Schizochytrium species atcC PTA-9695 (SEQ ID NO: 1), Schizochytrium species ATcc pta-9695 (SEQ ID NO: 3), Schizochytrium strain ATCC PTA-9695 (SEQ ID NO: 5), Thraustochytrium strain ATCCPTA-l〇212Pi^7 (SEQ ID NO: 68 or sequence identification No.: 12〇), multi-core of Thraustochytrium strain ATCC PTA-10212 (sequence identification number: 70 or sequence identification number: 121), Thraustochytrium strain ATCCPTA_10212PFw (sequence identification number: 72 or sequence identification number: 122) a sequence of a nucleoside, and a combination thereof, wherein the polynucleotides A polypeptide comprising one or more PUFA synthase activities. The PUFA synthase activity is associated with one or more domains of each synthetase polypeptide, wherein the domains are capable of conservation structure or function by Sexual elements (according to their homology to known elements) are identified, and may also be identified by their specific biochemical activity. See, for example, U.S. Patent No. 7,217,856, incorporated herein in its entirety. As a reference herein, examples of the PUFA synthetase include: the field of β-ketothiol-ACP synthase (KS) in Pfalp, the domain of propylenediyl-CoA: ACP thiol transferase (MAT), 醯The base carrier protein (ACP) domain, the ketoreductase 33 201038734 (KR) domain, and the β-radio-based Si-ACP dehydratase (DH) domain; the KS domain, the chain length factor (CLF) domain, and the thiol group in pfa2p The field of transferase (AT), and the dilute-ACP reductase (ER) domain; and the DH domain and ER domain within pfa3p. With β-ketothiol-ACP synthase (KS) biological activity ( Functionality) One of the fields of polypeptides or a polypeptide has previously been shown to be able to The initial step of the extended reaction cycle of the fatty acid. The term "β-ketodecyl-ACP synthase" has been combined with the term "3-ketodecyl-ACP synthase", "β-ketothiol -ACP synthase ", and " keto thiol, ACP synthase, are used interchangeably. In other systems, 'the activity of the thiol group linked to KS by a thioester bond has been shown to be extended. One of the cysteine residues at the position, as well as the thiol-ks system, condenses with the propylenediyl-ACP to form a keto fluorenyl group - Acp, c 〇 2 and unbound ("free" KS. In such systems, it has been shown that KS possesses greater receptor specificity than other polypeptides in the reaction cycle. The polypeptide (or domain of the polypeptide) can be readily identified as belonging to the KS family by homology to known KS sequences. One of the fields of poly(diphenyl)-CoA: ACP thiol transferase (MAT) activity Polymorphism or a polypeptide has previously been shown to be capable of transferring a propylenediyl moiety to ACP from propylenediyl-CoA. The term "propionyl-CoA: ACP thiol transferase" has been used interchangeably with "propionyl thiol transferase." In addition to the active site element (GxSxG), it has been shown that MATs have an extension. Elements (R and Q amino acids at key positions). Polypeptides (or domains of polypeptides) can be easily identified by their homology to known mat sequences and their extended elemental structures. For the family of mats. 34 201038734 The field of polypeptides or polypeptides having thiol-containing carrier protein (ACP) activity has previously been shown to act as a carrier of a growing fatty thiol chain, which is via a thioester bond. A cofactor that is linked to a covalent bond. ACPs are typically from about 80 to about 1 amino acid long and have been shown to transfer the phytosanban moiety on the CoA (ph〇Sph〇pantetheinyl moiety) ) to a highly conserved serine residue on ACP and converted from an inactive incomplete (apo)-form to a functionally complete (h〇i〇)_ form. It has also been shown to be based on phosphoric acid. Thioester bond of one of the free ends of the panthenol moiety Linked to ACPs. The presence of changes in active positional elements (LGIDS*) has also been identified as a feature of ACP. The functionality of this active position of serine acid (S*) has been demonstrated in a bacterial PUFA synthase (Jiang) Et al., j. Am. Chem. Soc. 130:6:336-7 (2〇〇8)). A polypeptide (or domain of a polypeptide) can be labeled by radioactive panthenol and by known The sequence homology of ACPs is easily identified as belonging to the ACP family. The field of polypeptides or polypeptides having dehydrase or dehydratase (DH) activity has previously been shown to be capable of catalyzing A dehydration reaction. Reference to DH activity typically refers to FabA-like β-hydroxyindenyl-ACP dehydratase biological activity. FabA-like β-hydroxyindenyl-ACP dehydratase biological activity will be derived from β-ketothiol -ACP removes Η0Η and initially creates a trans double bond on the carbon chain. The term "FabA-like β-hydroxyindenyl-ACP dehydratase" has been similar to the term "FabA-like β-hydroxyl醯-ACP dehydratase · ', πβ-hydroxy thiol-ACP dehydratase " and "dehydratase" exchange . The DH field of the PUFA synthase system has previously been shown to exhibit homologous bacterial DH enzymes associated with the FAS system (as opposed to the DH field of other PKS systems). See, for example, U.S. Patent No. 7,217,856, the disclosure of which is incorporated herein by reference. The subgroup of bacteria DH, which is similar to DH, possesses cis-trans isomerase activity (Heath et al., pp. 6丨〇1·Chem., 271, 27795 (1996)). Depending on the homology to the FabA-like DH protein, one or all of the DH domain of the PUFA synthase system can be responsible for the insertion of a cis double bond within the PUFA synthase product. A polypeptide or domain can also have non-FabA-like DH activity, or non-FabA-like β-hydroxyindenyl-ACP dehydratase (DH) activity. More specifically, a conserved active positional element of about 13 amino acids long has been previously identified in the field of PUFA synthase DH: LxxHxxxGxxxxP (the L position in this element may also be I). See, for example, U.S. Patent No. 7,217,856, and DonadioS, KatzL., Gene 111(1): 51-60 (1992), each of which is incorporated herein by reference in its entirety. This conservation factor is found in similar regions of all known PUFA synthase sequences and may be the cause of non-FabA-like dehydration. The field of a polypeptide or a polypeptide having /?-ketohydrazino-ACP reductase (KR) activity has previously been shown to be capable of catalyzing the pyridine-nucleotide-dependent type of ACP in the 3-ketoguanidino form. Restore effect. The term "wan-ketothiol-ACP reductase" has been combined with the term "ketone reductase", "3-ketothiol-ACP reductase', and "keto-mercapto ACP reductase "Interchangeable use. It has been determined that KR function in other systems involves the first reduction step of fatty acid re-synthesis to extend the cycle. The polypeptide (or domain of the polypeptide) can be homologous to the sequence of the known PUFA synthase KRs. It is readily identified as belonging to the KR family. 36 201038734 The field of polypeptides or polypeptides having one of chain length factor (CLF) activities has previously been defined as one or more of the following activities or properties: 1) It can determine the number of prolonged cycles and thus the chain length of the final product, (2) it has homology to KS 'but lacks KS active position cysteine, (3) it can be dimerized with KS (4) It can provide the initial test base to be extended, or (5) it can degrade the malonate (which is a propylenediamine-ACP), thus forming a transferable position to the KS active site. Acetate group and can be used as a starting and extending (condensation) reaction Priming, the molecule. The CLF domain is found in all recently identified PUFA synthase systems, and in each case it is found to be part of a multi-domain protein. The polypeptide (or the domain of the polypeptide) can be The sequence homology with the known PUFA synthase CLFs is easy. It is identified as belonging to the CLF family. The field of a polypeptide or a polypeptide having a thiotransferase (AT) activity has previously been defined as having One or more of the following activities or characteristics: (1) its ability to transfer the fat sulfhydryl group from the ACP domain to water (ie, thioester chymase), releasing the fatty sulfhydryl group as a free fatty acid, (2) It can transfer a fatty thiol group to an acceptor, such as CoA, (3) it can transfer a variety of sulfhydryl groups in the ACP domain, or (4) it can transfer fat thiol groups. To a lipophilic acceptor molecule (for example, to lys〇ph〇sphadic acid). The polypeptide (or domain of the polypeptide) can be easily homologous to the known pUFA synthetase ATs. It was identified as belonging to the AT family. Has a dilute sulfhydryl-ACP The field of one of the pro-enzyme (ER) biological activities of a polypeptide or a polypeptide has previously been shown to reduce the fatty sulfhydryl-trans double bond on ACP (introduced by DH activity)' resulting in associated carbon saturation puFA synthesis Enzyme 37 201038734 The field of the ruler has previously been shown to be homologous to the ER enzyme family (Heath et al, Nature 406: 145-146 (2000), incorporated herein by reference in its entirety), An ER homolog has been shown to act as an exo-alkali-ACP reductase in vitro (Bumpus et al., VIII „1.〇1抓·

Soc.，130: 11614-11616 (2008)’以其之整體併入本文中作為 參考資料）。術語”烯醯基-ACP還原酶"已經與，，烯醯基還原 酶"、"烯醯基ACP-還原酶"’及"烯酸基醯基—acp還原酶” 互相父換使用。多狀（或多狀的領域）能藉由與已知之PUfa 合成酶ERs之序列同源性而容易地被鑑定出為屬於該ER家 族。 於一些具體例中’本發明係針對核酸分子，其包含與 (序列辨識編號：1、序列辨識編號：68，或序列辨識 編號：120)中的多核苷酸序列有至少80%的同一性之多核苦 酸序列’其係編碼一或更多個PUFA合成酶領域。於一此具 體例中，該核酸分子包含與(序列辨識編號：1、序列 辨識編號：68，或序列辨識編號：120)中的多核苦酸序列 有至少80%的同一性之多核苷酸序列，其係編碼—或更多 個PUFA合成酶領域，例如：KS領域(序列辨識編號：7或序 列辨識編號：74)，MAT領域(序列辨識編號：9或序列辨識 編號：76)，ACP領域(例如：序列辨識編號：13、15、17、 19、21、23、80、82、84、86、88、90、92、94、96^98Soc., 130: 11614-11616 (2008) is incorporated herein by reference in its entirety. The term "alkenyl-ACP reductase" has been associated with, olefinic reductase ", olefinic ACP-reductase " &" enoylthiol-acp reductase Change to use. Polymorphic (or polymorphic domains) can be readily identified as belonging to the ER family by sequence homology to known PUfa synthetase ERs. In some embodiments, the invention is directed to a nucleic acid molecule comprising at least 80% identity to a polynucleotide sequence in (SEQ ID NO: 1, Sequence ID: 68, or Sequence ID: 120). The polynucleic acid sequence 'is encoded in the field of one or more PUFA synthetases. In one embodiment, the nucleic acid molecule comprises a polynucleotide sequence that is at least 80% identical to the polynucleotide sequence (SEQ ID NO: 1, Sequence ID: 68, or Sequence ID: 120) , its coding - or more PUFA synthetase fields, for example: KS field (sequence identification number: 7 or sequence identification number: 74), MAT field (sequence identification number: 9 or sequence identification number: 76), ACP field (Example: Sequence Identification Number: 13, 15, 17, 19, 21, 23, 80, 82, 84, 86, 88, 90, 92, 94, 96^98

的任何一者），2個或更多個ACP領域的組合，例如：2 ' 3、 4、5、6、7、8、9或1 〇個ACP領域，包括串聯領域(序列辨 識編號：11或序列辨識編號：78，以及其等之部分），KR 38 201038734 領域(序列辨識編號：25或序列辨識編號：1〇〇)，DH領域(序 列辨識編號：27或序列辨識編號：118)，以及其等之組合。 於一些具體例中，該核酸分子包含2個或更多個多核苷酸序 列’其中該至少2個或更多個多核苷酸序列的各個係與户柯/ (序列辨識編號：1、序列辨識編號：68，或序列辨識編號. 120)中的多核苦酸序列有80%的同一性，其係編碼一或更多 個PUFA合成酶領域。於一些具體例中，該至少2個或更多 0 個多核苷酸序列係與序列辨識編號：卜序列辨識編號：68， 或序列辨識編號：120中相同的多核苷酸序列有8〇%的同— 性，其係編碼一或更多個PUFA合成酶領域。於一些具體例 中’該至少2個或更多個多核苷酸序列係與序列辨識編號： 1、序列辨識編號：68，或序列辨識編號：120中不同的多 核苷酸序列有80%的同一性，其係各編碼一或更多個PUFA 合成酶領域。於一些具體例中，該至少2個或更多個多核苷 酸序列係與序列辨識編號：1、序列辨識編號：68，或序列 Q 辨識編號：120中不同的多核苷酸序列有80%的同一性，其 中當比較序列辨識編號：1、序列辨識編號：68，或序列辨 識編號：120中的對應序列的等級時，該至少2個或更多個 多核苷酸序列係位於該核酸分子中之相同的等級或不同的 等級。於一些具體例中，該至少2個或更多個多核苷酸序列 的各個係與户柯八序列辨識編號：1、序列辨識編號：68， 或序列辨識編號：120)中的多核苷酸序列有80%的同一性， 其係編碼一或更多個PUFA合成酶領域，例如：KS領域(序 列辨識編號：7或序列辨識編號：74)，MAT領域(序列辨識 39 201038734 編號：9或序列辨識編號：76) ’ ACP領域(例如：序列辨識 編號：13、15、17、19、21、23、80、82、84、86、88、 90、92、94、96或98 的任何一者），2、3、4、5、6、7、8、 9或10個ACP領域的組合，包括_聯領域(序列辨識編號：U 或序列辨識編號：78，以及其等之部分），KR領域(序列辨 識編號：25或序列辨識編號：100) ’ DH領域(序列辨識編號： 27或序列辨識編號：118) ’以及其等之組合。於一此具體 例中，該核酸分子包含户柯/(序列辨識編號：1、序列辨識 編说.68 ’或序列辨識編说.120)中的·一或更多個多核苦· 酸序列’其包括任何個別的領域之一或更多個副本與任何 其他個別的領域之一或更多個副本的組合。 於一些具體例中，本發明係針對核酸分子，其包含與 7>柯2(序列辨識編號：3、序列辨識編號：7〇 ,或序列辨識 編號：121)中的多核苷酸序列有至少8〇〇/0的同一性之多核苦 酸序列’其係編碼一或更多個PUFA合成酶領域。於一些具 體例中，該核酸分子包含與PE42(序列辨識編號：3、序列 辨識編號：70，或序列辨識編號：121)中的多核苷酸序列 有至少80%的同一性之多核苷酸序列，其係編碼—或更多 個PUFA合成酶領域，例如：KS領域(序列辨識編號：29或 序列辨識編號：102)，CLF領域(序列辨識編號：31或序列 辨識編號：104)，AT領域(序列辨識編號：33或序列辨識編 號：106)，ER領域(序列辨識編號：35或序列辨識編號： 108)，以及其等之組合。於一些具體例中，該核酸分子包 含2個或更多個多核苷酸序列，其中該至少2個或更多個多 201038734 核苷酸序列的各個係與户柯2(序列辨識編號^ j、序列辨識 、’爲唬· 70 ’或序列辨識編號：121)中的多核苷酸序列有8〇〇/ 的同一性，其係編碼一或更多個PUFA合成酶領域。於此 具體例中，縣少2個或更多個多核賴相係與序列辨^ 編號：3、序列辨識編號：7〇，或序列辨識編號：a〗中相 同的多核苷酸序列有80%的同一性，其係編碼—或更多個 PUFA合成酶領域。於一些具體例中，該至少]個或更=個 多核苷酸序列係與序列辨識編號：3、序列辨識編號：川， 或序列辨識編號：121中不同的多核苷酸序列有8〇%的同一 性，其係各編碼一或更多個PUFA合成酶領域。於一些具體 例中，該至少2個或更多個多核苷酸序列係與序列辨識編 號：3、序列辨識編號：70，或序列辨識編號：12丨中不同 的多核賊序列有80%的同-性，其中當比較序列辨識編 號：3、序列辨識編號：70,或序列辨識編號：i2i中的對 應序列的等級時’該至少2個或更多個多核苷酸序列係位於 該核酸分子中之相同的等級或不同的等級。於一些具體例 中，該至少2個或更多個多核苷酸序列的各個係與户柯2 (序 列辨識編號.3、序列辨識編號：7〇，或序列辨識編號：121) 中的多核皆酸序列有80%的同一性，其係編碼一或更多個 PUFA合成酶領域，例如：KS領域(序列辨識編號：29或序 列辨識編號.102)，CLF領域(序列辨識編號：31或序列辨 識編號：1〇4)’AT領域(序列辨識編號：33或序列辨識編號： 106)，ER領域(序列辨識編號：35或序列辨識編號：108)， 以及其等之組合。於一些具體例中，該核酸分子包含p柯2 201038734 (序列辨識編號：3、序列辨識編號：％ 次序列辨識編號： ⑵）中的或更夕個夕核倾序列，其柄碼 疆合成酶領域，其包括任何個別的領域二夕個 本與任何其他_的領域之—或更多個副本的組合。夕固Μ 於-些具體例中，本發明係針對核酸分子，呈 削3 (序列辨識編號：5、序列辨識編號：72，或序列辨識 編號：12 2)巾料核料序财至少8 G %的同—性之多核芽 酸序列’其係編碼-或更多個PUFA合成酶領域。於一些具 體例中，該核酸分子包含與则3 (序列辨識編號：5、序; 辨識編號：72，或序列辨識編號：122)中的多核苷酸序列 有至少80%的同-性之多核皆酸序歹，卜其係編碼—或更多 個PUFA合成酶領域，例如：DH領域(例如：序列辨識編號： 37，序列辨識編號：39，序列辨識編號：11〇 ,或序列辨識 編號：112)，ER領域(序列辨識編號：41或序列辨識編號： 114)，以及其4之組合。於一些具體例中，該核酸分子包 含2個或更多個多核苷酸序列，其中該至少2個或更多個多 核苷酸序列的各個係與/>柯3 (序列辨識編號：5、序列辨識 編號：72 ’或序列辨識編號：122)中的多核苷酸序列有8〇% 的同一性，其係編碼一或更多個PUFA合成酶領域。於一些 具體例中，該至少2個或更多個多核苷酸序列係與序列辨識 編號：5、序列辨識編號·· 72，或序列辨識編號：122中相 同的多核苷酸序列有80%的同一性，其係編碼—或更多個 PUFA合成酶領域。於一些具體例中，該至少2個或更多個 多核苷酸序列係與序列辨識編號：5、序列辨識編號：72， 42 201038734 或序列辨識編號：122中不同的多核苷酸序列有80%的同一 性，其係各編碼一或更多個PUFA合成酶領域。於一些具體 例中，該至少2個或更多個多核苷酸序列係與序列辨識編 號：5、序列辨識編號：72，或序列辨識編號：122中不同 的多核普酸序列有80%的同一性，其中當比較序列辨識編 號：5、序列辨識編號：72，或序列辨識編號：122中的對 應序列的等級時’該至少2個或更多個多核苷酸序列係位於 該核酸分子中之相同的等級或不同的等級。於一些具體例 中，該至少2個或更多個多核苷酸序列的各個係與户抑3 (序 列辨識編號：5、序列辨識編號：72，或序列辨識編號：122) 中的多核苷酸序列有80%的同一性，其係編碼一或更多個 PUFA合成酶領域，例如：DH領域(例如：序列辨識編號： 37，序列辨識編號：39 ’序列辨識編號：11〇，或序列辨識 編號：112)，ER領域(序列辨識編號：41或序列辨識編號： 114)，以及其等之組合。於一些具體例中，該核酸分子包 含户柯3 (序列辨識編號：5、序列辨識編號：72，或序列辨 識編號：122)中的一或更多個多核苷酸序列，其係編碼一 或更多個PUFA合成扭領域，其包括任何個別的領域之一或 更多個副本與任何其他個別的領域之一或更多個副本的組 合。Any one of 2, or more combinations of ACP fields, for example: 2 ' 3, 4, 5, 6, 7, 8, 9 or 1 ACP fields, including tandem fields (sequence identification number: 11 Or sequence identification number: 78, and its parts), KR 38 201038734 field (sequence identification number: 25 or sequence identification number: 1〇〇), DH field (sequence identification number: 27 or sequence identification number: 118), And a combination of them. In some embodiments, the nucleic acid molecule comprises two or more polynucleotide sequences 'where each of the at least two or more polynucleotide sequences is SEQ ID NO: 1. Sequence identification The polynucleotide sequence in Accession No.: 68, or Sequence Identification Number. 120) is 80% identical and encodes one or more of the PUFA synthetase domains. In some embodiments, the at least two or more polynucleotide sequences are SEQ ID NO: 68, or the same polynucleotide sequence in Sequence Identification Number: 120 is 8%. Synonymous, which encodes one or more of the PUFA synthetase domains. In some embodiments, the at least two or more polynucleotide sequences are 80% identical to the sequence number of the sequence identification number: 1, the sequence identification number: 68, or the sequence identification number: 120. Sexuality, each of which encodes one or more of the PUFA synthetase domains. In some embodiments, the at least two or more polynucleotide sequences are 80% identical to the sequence identification number: 1, the sequence identification number: 68, or the sequence Q identification number: 120 different polynucleotide sequences. Identity, wherein when comparing the sequence identification number: 1, the sequence identification number: 68, or the sequence of the corresponding sequence in the sequence identification number: 120, the at least two or more polynucleotide sequences are located in the nucleic acid molecule The same level or different levels. In some embodiments, the polynucleotide sequence of each of the at least two or more polynucleotide sequences is SEQ ID NO: 1, sequence identification number: 68, or sequence identification number: 120) 80% identity, which encodes one or more PUFA synthetase domains, eg KS domain (sequence identification number: 7 or sequence identification number: 74), MAT domain (sequence identification 39 201038734 number: 9 or sequence Identification number: 76) 'ACP field (eg sequence identification number: any of 13, 15, 17, 19, 21, 23, 80, 82, 84, 86, 88, 90, 92, 94, 96 or 98) ), 2, 3, 4, 5, 6, 7, 8, 9 or 10 combinations of ACP fields, including _ associated fields (sequence identification number: U or sequence identification number: 78, and parts thereof), KR Field (sequence identification number: 25 or sequence identification number: 100) 'DH field (sequence identification number: 27 or sequence identification number: 118) 'and combinations thereof. In one embodiment, the nucleic acid molecule comprises one or more polynuclear acid sequences in the family/(SEQ ID NO: 1, Sequence Identification Editor. 68 ' or Sequence Identification Editor. 120) It includes a combination of one or more copies of any individual field with one or more copies of any other individual field. In some embodiments, the invention is directed to a nucleic acid molecule comprising at least 8 polynucleotide sequences in 7> Ke 2 (SEQ ID NO: 3, Sequence ID: 7〇, or Sequence ID: 121) The polynucleic acid sequence of 〇〇/0 identity is encoded in the field of one or more PUFA synthetases. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence that is at least 80% identical to the polynucleotide sequence in PE42 (SEQ ID NO: 3, SEQ ID NO: 70, or SEQ ID NO: 121) , in the field of coding - or more PUFA synthetases, for example: KS domain (sequence identification number: 29 or sequence identification number: 102), CLF domain (sequence identification number: 31 or sequence identification number: 104), AT field (Sequence Identification Number: 33 or Sequence Identification Number: 106), ER field (Sequence Identification Number: 35 or Sequence Identification Number: 108), and combinations thereof. In some embodiments, the nucleic acid molecule comprises two or more polynucleotide sequences, wherein each of the at least two or more multiple 201038734 nucleotide sequences and the family 2 (SEQ ID NO: The polynucleotide sequence in sequence identification, 'Yes 70' or Sequence ID: 121) has 8 〇〇 identity, which encodes one or more PUFA synthetase domains. In this specific example, the county has less than two or more multi-nucleolioid phases and the sequence identification number: 3, the sequence identification number: 7〇, or the same polynucleotide sequence in the sequence identification number: a is 80%. Identity, which is encoded in the field of more than one PUFA synthetase. In some embodiments, the at least one or more polynucleotide sequence lines and the sequence identification number: 3, the sequence identification number: Chuan, or the sequence identification number: 121 different polynucleotide sequences have 8〇% Identity, which is in the field of encoding one or more PUFA synthetases. In some embodiments, the at least two or more polynucleotide sequences are 80% identical to the sequence identification number: 3, the sequence identification number: 70, or the sequence identification number: 12 多 different multi-core thief sequences. - sex, wherein when comparing the sequence identification number: 3, the sequence identification number: 70, or the sequence identification number: the level of the corresponding sequence in i2i 'the at least 2 or more polynucleotide sequences are located in the nucleic acid molecule The same level or different levels. In some embodiments, each of the at least two or more polynucleotide sequences is a multi-nucleus in the family 2 (sequence identification number 3., sequence identification number: 7〇, or sequence identification number: 121) The acid sequence is 80% identical and encodes one or more PUFA synthetase domains, for example: KS domain (SEQ ID NO: 29 or sequence identification number. 102), CLF domain (SEQ ID NO: 31 or sequence) Identification number: 1〇4) 'AT field (sequence identification number: 33 or sequence identification number: 106), ER field (sequence identification number: 35 or sequence identification number: 108), and combinations thereof. In some embodiments, the nucleic acid molecule comprises p-C 2 201038734 (SEQ ID NO: 3, SEQ ID NO: % SEQ ID NO: (2)) The field, which includes any combination of individual fields and any other field, or a combination of multiple copies. In some specific examples, the present invention is directed to a nucleic acid molecule, which is cut 3 (sequence identification number: 5, sequence identification number: 72, or sequence identification number: 12 2). % of the homozygous polynuclear acid sequence 'the line encodes - or more than one PUFA synthetase field. In some embodiments, the nucleic acid molecule comprises at least 80% homozygous multinuclear with a polynucleotide sequence in the sequence 3 (SEQ ID NO: 5, Sequence; ID: 72, or Sequence ID: 122) All acid sequence, encoding or more - in the field of PUFA synthetase, for example: DH domain (eg: sequence identification number: 37, sequence identification number: 39, sequence identification number: 11〇, or sequence identification number: 112), ER field (sequence identification number: 41 or sequence identification number: 114), and a combination of 4. In some embodiments, the nucleic acid molecule comprises two or more polynucleotide sequences, wherein each of the at least two or more polynucleotide sequences and /> Ke 3 (sequence identification number: 5, The polynucleotide sequence in Sequence Identification Number: 72' or Sequence Identification Number: 122 has 8% identity identity and encodes one or more PUFA synthetase domains. In some embodiments, the at least two or more polynucleotide sequences are 80% identical to the same polynucleotide sequence number: 5, sequence identification number 72, or sequence identification number: 122. Identity, which is encoded in the field of more than one PUFA synthase. In some embodiments, the at least two or more polynucleotide sequences are 80% different from the sequence number of the sequence identification number: 5, sequence identification number: 72, 42 201038734 or sequence identification number: 122. Identity, which is in the field of encoding one or more PUFA synthetases. In some embodiments, the at least two or more polynucleotide sequences are 80% identical to the sequence identification number: 5, sequence identification number: 72, or different polynucleotide sequences in sequence identification number: 122. Sex, wherein when comparing the sequence identification number: 5, the sequence identification number: 72, or the sequence of the corresponding sequence in the sequence identification number: 122, the at least two or more polynucleotide sequences are located in the nucleic acid molecule. The same level or different levels. In some embodiments, the polynucleotides of each of the at least two or more polynucleotide sequences are SEQ ID NO: 5, SEQ ID NO: 72, or SEQ ID NO: 122. The sequence is 80% identical and encodes one or more of the PUFA synthetase domains, eg, the DH domain (eg, sequence ID: 37, sequence ID: 39 'sequence ID: 11〇, or sequence identification) No.: 112), ER field (sequence identification number: 41 or sequence identification number: 114), and combinations thereof. In some embodiments, the nucleic acid molecule comprises one or more polynucleotide sequences in C. 3 (SEQ ID NO: 5, SEQ ID NO: 72, or SEQ ID NO: 122), which encodes one or More PUFA Synthetic Twisted Fields, which include a combination of one or more copies of any individual field with one or more copies of any other individual field.

於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編號：1、序列辨識編號：68，或序列辨識編號： 120有至少80%的同一性之多核苷酸序列，其中該多核苷酸 序列編碼一多肽，其包含選自於以下所構成的群組之puFA 43 201038734 合成酶活性：KS活性、MAT活性、ACP活性、KR活性、 DH活性、以及其等之組合。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編號：7或序列辨識編號：74有至少80%的同一 性之多核苷酸序列，其中該多核苷酸序列編碼一多肽，其 包含KS活性。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編號：9或序列辨識編號：76有至少80%的同一 性之多核普酸序列，其中該多核普酸序列編碼一多肽，其 包含MAT活性。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編號：13、15、17、19、21、23、80、82、84、 86、88、90、92、94、96或98的任一者有至少80%的同一 性之多核苷酸序列，其中該多核苷酸序列編碼一多肽，其 包含ACP活性。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編號：11或序列辨識編號：78有至少80%的同 一性之多核苦酸序列，其中該多核苦酸序列編碼一多肽， 其包含ACP活性。 於一些具體例中，該核酸分子包含與序列辨識編號： 11中的多核苷酸序列有至少8 0%的同一性之多核苷酸序 列，其編碼1、2、3、4、5或6個ACP領域，其中該多核苷 酸序列編碼一多肽，其包含與一或更多個ACP領域關聯的 ACP活性。序列辨識編號：13、15、17、19、21，與23為 44 201038734 代表性多核苷酸序列，其各編碼序列辨識編號：u内的單 一 ACP領域。 於一些具體例中，該核酸分子包含與序列辨識編號： 78中的多核苷酸序列有至少8〇%的同一性之多核苷酸序 列’其編碼1、2、3、4、5、6、7、8、9或10個ACP領域， 其中該多核苷酸序列編碼一多肽，其包含與一或更多個 ACP領域關聯的ACP活性。序列辨識編號：8〇、82、84、 〇 86、88、90、92、94、96與卯為代表性多核苷酸序列，其 各編碼序列辨識編號：78内的單一ACP領域。 於一些具體例中，本發明係針對一核酸分子，其包含 - 與序列辨識編號：25或序列辨識編號：ι〇〇有至少8〇%的同 . —性之多核苷酸序列，其中該多核苷酸序列編碼一多肽， 其包含KR活性。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編號：27或序列辨識編號：U8有至少8〇%的同 〇 性之多核苷酸序列，其中該多核苷酸序列編碼一多肽， 其包含DH活性。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編號3、序列辨識編號：70,或序列辨識編號： 121有至少8〇。/〇的同一性之多核苷酸序列，其中該多核苷酸 序列編碼一多肽，其包含選自於以下所構成的群組之p UFA 合成酶活性：KS活性、CLF活性、AT活性、ER活性、以及 其等之組合。 於一些具體例中，本發明係針對—核酸分子，其包含 45 201038734 與序列辨識編號：29或序列辨識編號：102有至少80%的同 一性之多核苦酸序列’其中該多核苦酸序列編碼一多肽， 其包含KS活性。 於一些具體例中’本發明係針對一核酸分子，其包含 與序列辨識編號：31或序列辨識編號：104有至少80%的同 一性之多核苷酸序列，其中該多核苷酸序列編碼一多狀， 其包含CLF活性。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編號：33或序列辨識編號：1〇6有至少80%的同 一性之多核苷酸序列，其中該多核苷酸序列編碼—多肽， 其包含AT活性。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編號：35或序列辨識編號：1〇8有至少8〇%的同 一性之多核苷酸序列，其中該多核苷酸序列編碼一多肽， 其包含ER活性。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編號：5、序列辨識編號：72,或序列辨識編號. 122有至少80%的同一性之多核苷酸序列’其中該多核苷酸 序列編碼一多肽’其包含選自於以下所構成的群組之puFA 合成酶活性：DH活性、ER活性、以及其等之組合。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編"5虎.37有至少8〇%的同一性之多核;y：酸序 列，其中該多核苷酸序列編碼一多肽，其包含〇1{活性。 於一些具體例中，本發明係針對一核酸分子，其包含 46 201038734 與序列辨識編號：39有至少800/。的同一性之多核苷酸序 列’其中该多核有:酸序列編瑪一多狀’其包含DH活性。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編婕：110有系少80%的同一性之多核苷酸序 列，其中δ亥多核苷酸序列編瑪一多肽，其包含DH活性。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編說：丨12有炱少80%的同一性之多核苷酸序 列’其中§亥多核芽酸序列編碼一多肽’其包含DH活性。 於一些具體例中，本發明係針對一核酸分子，其包含 與序列辨識編號：41或序列辨識編號：114有至少80%的同 一性之多核苷酸序列，其中該多核苷酸序列編碼一多肽， 其包含ER活性。 本發明係針姆經單離的核酸分子，其包含編碼多肽之 多核苷酸序列，其中該等多肽包含與Pfalp (序列辨識編 號：2或序列辨識編號：69)、Pfa2P (序列辨識編號：4或序 列辨識編號：71)，或是Pfa3p (序列辨識編號：6或序列辨 識編號：73)之胺基酸序列有至少80%的同一性之胺基酸序 列，其中該多核苷酸編碼包含一或更多PUFA合成酶活性之 多肽。 本發明係針對核酸分子’其包含編碼一多肽之多核苷 酸序列，其中該多肽包含與本發明的PUFA合成酶之一或更 多個PUFA合成酶領域的胺基酸序列有至少80%的同一性之 胺基酸序列。 於一些具體例中’本發明係針對核酸分子’其包含編 47 201038734 碼-多肽之多核皆酸序列’其中該多狀包含與pfai(序列辨 識編號.2或序列辨識編號：69)中的胺基酸序列有至少祕 的同-性之胺基酸序列，其包含—或更多個puFA合成酶領 域。於-些具體例中’該多肽包含與pfalp (序列辨識編號： 2或序列辨識編號：的)中的胺基酸序列有至少議的同一性 之胺基酸序列，其包含—或更多個pUFA合成酶領域，例 如：KS領域(序列辨識編號：8或序列辨識編號：75)，MAT 領域(序列辨識編號：10或序列辨識編號：77)，ACp領域(例 如：序列辨識編號：14、16、18、20、22、24、81、83、 85、87 ' 89、91、93、95、97或99的任何一者），2個或更 多個ACP領域的組合，例如：2、3、4、5、6、7、8、9或 10個ACP領域，包括串聯領域(序列辨識編號：12或序列辨 識編號：79，以及其等之部分），KR領域(序列辨識編號： 26或序列辨識編號：i〇1)，DH領域(序列辨識編號：28或序 列辨識編號：119)，以及其等之組合。於一些具體例中， 該多肽包含2個或更多個胺基酸序列，其中該至少2個戍更 多個胺基酸序列的各個係與Pfalp(;序列辨識編號：2或序列 辨識編號：69)中的胺基酸序列有80%的同一性，其包含— 或更多個PUFA合成酶領域。於一些具體例中，該至少2個 或更多個胺基酸序列與P fa 1 p (序列辨識編號：2或序列辨識 編號：69)中相同的胺基酸序列有80%的同一性，其包人 或更多個PUFA合成酶領域。於一些具體例中，該至少2個 或更多個胺基酸序列與P fa 1 p (序列辨識編號：2或序列辨識 編號：69)中不同的胺基酸序列有8〇〇/。的同一性，其各勺人 48 201038734 一或更多個PUFA合成酶領域。於一些具體例中，該至少2 個或更多個胺基酸序列與Pfalp (序列辨識編號：2或序列辨 識編號：69)中不同的胺基酸序列有80%的同—性，其中春 比較Pfalp (序列辨識編號：2或序列辨識編號：69)令的對應 領域的專級日寸’ s亥至少2個或更多個胺基酸序列係位於該多 肽中之相同的等級或不同的等級。於一些具體例中，該至 少2個或更多個胺基酸序列與Pfalp (序列辨識編號：2或序 0 列辨識編號：69)中的胺基酸序列有80%的同一性，其包含 一或更多個PUFA合成酶領域，例如：KS領域(序列辨識編 號：8或序列辨識編號：75)，MAT領域(序列辨識編號：1〇 - 或序列辨識編號：77)，ACP領域(例如：序列辨識編號：14、 16、18、20、22、24、8卜 83、85、87、89、91、93、95、 97或99的任何一者）’ 2、3、4、5、6、7、8、9或 1〇個ACP 領域的組合，包括串聯領域(序列辨識編號：12或序列辨識 編號：79，以及其等之部分），KR領域(序列辨識編號：％ Ο 或序列辨識編號：101)，DH領域(序列辨識編號：28或序列 辨識編號：119)，以及其等之組合。於一些具體例中，該 多肽包含Pfalp(序列辨識編號：2或序列辨識編號：69)中的 1個或更多個胺基酸序列，其包含一或更多個PUFA合成酶 領域’其包括任何個別的領域之一或更多個副本與任何其 他個別的領域之一或更多個副本的組合。 於—些具體例中，本發明係針對核酸分子，其包含編 碼一多肽之多核苷酸序列’其中該多肽包含與Pfa2p (序列 辨識編號：4或序列辨識編號：71)中的胺基酸序列有至少 49 201038734 的同n之胺基酸序列，其包含—或更多個puFA合成 酶員域於些具體例中，該多狀包含與跑如（序列辨識 、扁说4或序列辨識編號：7丨）中的胺基酸序列有至少⑽％的 同I·生之胺基酸序列，其包含一或更多個合成酶領 域例如，KS領域(序列辨識編號：3〇或序列辨識編號： 1〇3) ’ CLF領域(序列辨識編號·· 32或序列辨識編號：ι〇5)， AT錢(序列辨識編號：34或序列辨識編號：⑻），謝員域 (序列辨識編號：36或相: 109)，以及其等之組 口於些具體例中，該多肽包含2個或更多個胺基酸序 列’其中該至少2個或更多個胺基酸序列的各個係與 Pfa2p(序列辨識編號：4或序列辨識編號：中的胺基酸序 列有80%的同一性，其包含一或更多個pTJFA合成酶領域。 於一些具體例中’該至少2個或更多個胺基酸序列與pfa2p (序列辨識編號：4或序列辨識編號：71)中相同的胺基酸序 列有80%的同一性。於一些具體例中，該至少2個或更多個 胺基酸序列與Pfa2p (序列辨識編號：4或序列辨識編號：71) 中不同的胺基酸序列有80%的同一性，其各包含一或更多 個PUFA合成酶領域。於一些具體例中，該至少2個或更多 個胺基酸序列與Pfa2p(序列辨識編號：4或序列辨識編號： 71)中不同的胺基酸序列有80%的同一性，其中當比較Pfa2p (序列辨識編號：4或序列辨識編號：71)中的對應領域的等 級時，該至少2個或更多個胺基酸序列係位於該多肽中之相 同的等級或不同的等級。於一些具體例中’該至少2個或更 多個胺基酸序列與Pfa2p (序列辨識編號：4或序列辨識編 50 201038734 號：71)中的胺基酸序列有8〇%的同一性，其包含一或更多 個PUFA合成酶領域，例如：KS領域(序列辨識編號：3〇或 序列辨識編號.103)，CLF領域(序列辨識編號：32或序列 辨識編號：105)，AT領域(序列辨識編號：34或序列辨識編 號：107) ’ ER領域(序列辨識編號：36或序列辨識編號： 109)，以及其等之組合。於一些具體例中，該多肽包含pfa2p (序列辨識編5虎.4或序列辨識編號：71)中的1個或更多個胺 基酸序列，其包含一或更多個PUFA合成酶領域，其包括任 何個別的領域之一或更多個副本與任何其他個別的領域之 一或更多個副本的組合。 於一些具體例中，本發明係針對核酸分子，其包含編 碼-多肽之多核皆酸序列，其中該多肽包含與pfa3p中的胺 基酸序列(序列辨識編號：6或序列辨識編號：73)有至少8〇% 的同-性之胺基酸序列，其包含—或更多個pUFA合成酶領 域。於-些具體射，該多肽包含與Pfa3p中的胺基酸序列 (序列辨識編號：6或序列辨識編號：73)有至少8〇%的同一 性t胺ϋ含-或更多個PUFA合成酶領域，例 如：DH領域(例如：序列辨識編號：38、序列辨識編號：4〇、 序列辨識編號：in，或序列辨識編號：113)，ER領域(序 列辨識編號：42或序列辨識編號：ι15)，以及其等之組八。 於-些具體例中，該多肽包含2個或更多個胺基酸序列^ 中該至少2個或更多個胺基酸序列的各個係與pfa3p(序列辨 識編號：6或序列辨識編號：73)中的胺基酸序列有8〇%的同 -性，其包含-或更多個PUFA合成酶領域。於一些具體例 51 201038734In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence having a sequence identity number: 1, a sequence identification number: 68, or a sequence identification number: 120 having at least 80% identity, wherein The polynucleotide sequence encodes a polypeptide comprising puFA 43 201038734 synthetase activity selected from the group consisting of KS activity, MAT activity, ACP activity, KR activity, DH activity, and combinations thereof. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to Sequence ID: 7 or Sequence ID: 74, wherein the polynucleotide sequence encodes a plurality A peptide comprising KS activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a multi-nucleotide sequence having at least 80% identity to sequence identification number: 9 or sequence identification number: 76, wherein the polynucleotide sequence encodes more than one A peptide comprising MAT activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising and sequence identification numbers: 13, 15, 17, 19, 21, 23, 80, 82, 84, 86, 88, 90, 92, 94, 96 Or any of 98 having a polynucleotide sequence of at least 80% identity, wherein the polynucleotide sequence encodes a polypeptide comprising ACP activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 11 or sequence identification number: 78, wherein the polynucleotide sequence encodes a plurality of Peptide, which contains ACP activity. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence that is at least 80% identical to the polynucleotide sequence of Sequence Identification Number: 11, encoding 1, 2, 3, 4, 5 or 6 In the ACP field, wherein the polynucleotide sequence encodes a polypeptide comprising ACP activity associated with one or more ACP domains. Sequence identification numbers: 13, 15, 17, 19, 21, and 23 are 44 201038734 representative polynucleotide sequences, each of which has a single sequence identification number: u in the ACP field. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence having at least 8% identity with the polynucleotide sequence of Sequence Identification Number: 78, which encodes 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 ACP domains, wherein the polynucleotide sequence encodes a polypeptide comprising ACP activity associated with one or more ACP domains. Sequence identification numbers: 8〇, 82, 84, 〇 86, 88, 90, 92, 94, 96 and 卯 are representative polynucleotide sequences, each of which has a single ACP domain within the coding sequence number: 78. In some embodiments, the invention is directed to a nucleic acid molecule comprising - a polynucleotide sequence having a sequence identity number: 25 or a sequence identification number: ι 至少 having at least 8 %, wherein the polynucleotide The nucleotide sequence encodes a polypeptide comprising KR activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence having at least 8% homology to a sequence ID: 27 or a sequence number: U8, wherein the polynucleotide sequence is encoded A polypeptide comprising DH activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising at least 8 Å with sequence identification number 3, sequence identification number: 70, or sequence identification number: 121. a polynucleotide sequence of the identity of 〇, wherein the polynucleotide sequence encodes a polypeptide comprising p UFA synthase activity selected from the group consisting of KS activity, CLF activity, AT activity, ER Activity, and combinations thereof. In some embodiments, the invention is directed to a nucleic acid molecule comprising 45 201038734 a polynucleotide sequence having at least 80% identity to sequence identification number: 29 or sequence identification number: 102, wherein the polynucleotide sequence is encoded by the polynucleotide A polypeptide comprising KS activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 31 or sequence identification number: 104, wherein the polynucleotide sequence encodes a plurality of It contains CLF activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 33 or sequence identification number: 1〇6, wherein the polynucleotide sequence is encoded - a polypeptide comprising AT activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence having a sequence identity number: 35 or a sequence identification number: 1〇8 having at least 8% identity, wherein the polynucleotide sequence A polypeptide encoding an ER activity is encoded. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence having a sequence identity number: 5, a sequence identification number: 72, or a sequence identification number of 122 having at least 80% identity. The polynucleotide sequence encodes a polypeptide comprising a puFA synthase activity selected from the group consisting of DH activity, ER activity, and combinations thereof. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleus having at least 8% identity with the sequence recognition code "5 Tiger.37; y: an acid sequence, wherein the polynucleotide sequence encodes a A polypeptide comprising 〇1{activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising 46 201038734 and sequence identification number: 39 having at least 800/. The polynucleotide sequence of identity 'where the polynucleus has: the acid sequence is a polymorphous' which contains DH activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence that is 80% less identical to the sequence recognition codon: 110, wherein the delta polynucleotide sequence encodes a polypeptide, It contains DH activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence that is 80% identical to the sequence recognition: 丨12 has a nucleotide sequence encoding 'a polypeptide' It contains DH activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to Sequence ID: 41 or Sequence ID: 114, wherein the polynucleotide sequence encodes a plurality A peptide comprising ER activity. The present invention is a nucleic acid molecule which is isolated and comprises a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises Pfalp (SEQ ID NO: 2 or Sequence ID: 69), Pfa2P (SEQ ID NO: 4 Or the sequence identification number: 71), or the amino acid sequence of the amino acid sequence of Pfa3p (SEQ ID NO: 6 or Sequence ID: 73) having at least 80% identity, wherein the polynucleotide coding comprises a Or more polypeptides of PUFA synthase activity. The present invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% of an amino acid sequence in the field of one or more PUFA synthase enzymes of the PUFA synthase of the invention The amino acid sequence of identity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynuclear acid sequence of the 47 201038734 code-polypeptide, wherein the polymorph comprises an amine in pfai (SEQ ID NO: 2 or Sequence ID: 69) The acyl acid sequence has at least a secret homo-amino acid sequence comprising - or more puFA synthetase domains. In some embodiments, the polypeptide comprises an amino acid sequence at least having the identity of an amino acid sequence in pfalp (SEQ ID NO: 2 or Sequence ID:), which comprises - or more In the field of pUFA synthetase, for example: KS domain (sequence identification number: 8 or sequence identification number: 75), MAT domain (sequence identification number: 10 or sequence identification number: 77), ACp domain (eg sequence identification number: 14, 16, 18, 20, 22, 24, 81, 83, 85, 87 'any of 89, 91, 93, 95, 97 or 99), a combination of 2 or more ACP fields, for example: 2 3, 4, 5, 6, 7, 8, 9 or 10 ACP fields, including tandem fields (sequence identification number: 12 or sequence identification number: 79, and parts thereof), KR field (sequence identification number: 26 Or sequence identification number: i〇1), DH field (sequence identification number: 28 or sequence identification number: 119), and combinations thereof. In some embodiments, the polypeptide comprises two or more amino acid sequences, wherein each of the at least two 戍 more amino acid sequences is associated with Pfalp (SEQ ID NO: 2 or Sequence ID: The amino acid sequence in 69) is 80% identical and comprises - or more than one PUFA synthetase domain. In some embodiments, the at least two or more amino acid sequences are 80% identical to the same amino acid sequence in P fa 1 p (SEQ ID NO: 2 or SEQ ID NO: 69), It is in the field of human or more PUFA synthetases. In some embodiments, the at least two or more amino acid sequences differ from the amino acid sequence of P fa 1 p (SEQ ID NO: 2 or SEQ ID NO: 69) by 8 〇〇. The identity of each scoop person 48 201038734 One or more PUFA synthetase domains. In some embodiments, the at least two or more amino acid sequences are 80% homologous to the amino acid sequence different from Pfalp (SEQ ID NO: 2 or SEQ ID NO: 69), wherein Comparing Pfalp (Sequence Identification Number: 2 or Sequence Identification Number: 69) allows the corresponding field of the corresponding field to have at least 2 or more amino acid sequences located at the same level or different in the polypeptide. grade. In some embodiments, the at least two or more amino acid sequences are 80% identical to the amino acid sequence in Pfalp (SEQ ID NO: 2 or Sequence 0, ID: 69), which comprises One or more fields of PUFA synthetase, for example: KS domain (sequence identification number: 8 or sequence identification number: 75), MAT domain (sequence identification number: 1〇- or sequence identification number: 77), ACP domain (eg : Sequence identification number: any of 14, 14, 18, 20, 22, 24, 8 83, 85, 87, 89, 91, 93, 95, 97 or 99) 2, 3, 4, 5, 6, 7, 8, 9 or 1 combination of ACP fields, including tandem fields (sequence identification number: 12 or sequence identification number: 79, and parts thereof), KR field (sequence identification number: % Ο or sequence) Identification number: 101), DH field (sequence identification number: 28 or sequence identification number: 119), and combinations thereof. In some embodiments, the polypeptide comprises one or more amino acid sequences in Pfalp (SEQ ID NO: 2 or Sequence ID: 69) comprising one or more PUFA synthetase domains' A combination of one or more copies of any individual field with one or more copies of any other individual field. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide wherein the polypeptide comprises an amino acid in Pfa2p (SEQ ID NO: 4 or Sequence ID: 71) The sequence has at least 49 201038734 of the same n-amino acid sequence, which comprises - or more than one puFA synthase domain, in some specific examples, the polymorphism comprises and runs as (sequence identification, flat 4 or sequence identification number) The amino acid sequence of :7丨) has at least (10)% of the same amino acid sequence, which comprises one or more synthetic enzyme domains, for example, the KS domain (SEQ ID NO: 3〇 or sequence identification number : 1〇3) 'CLF field (sequence identification number··32 or sequence identification number: ι〇5), AT money (sequence identification number: 34 or sequence identification number: (8)), thank-you field (sequence identification number: 36) Or phase: 109), and combinations thereof, in some embodiments, the polypeptide comprises two or more amino acid sequences 'where each of the at least two or more amino acid sequences is associated with Pfa2p (Sequence identification number: 4 or sequence identification number: amino acid sequence in Listed as having 80% identity, comprising one or more domains of pTJFA synthetase. In some embodiments, the at least two or more amino acid sequences and pfa2p (SEQ ID NO: 4 or sequence identification number The same amino acid sequence in :71) is 80% identical. In some embodiments, the at least 2 or more amino acid sequences and Pfa2p (SEQ ID NO: 4 or Sequence ID: 71) The different amino acid sequences are 80% identical, each comprising one or more PUFA synthetase domains. In some embodiments, the at least two or more amino acid sequences and Pfa2p (sequence recognition) Number: 4 or sequence identification number: 71) 80% identity of different amino acid sequences, wherein when comparing the level of the corresponding field in Pfa2p (sequence identification number: 4 or sequence identification number: 71), At least 2 or more amino acid sequences are located at the same level or different levels in the polypeptide. In some embodiments, the at least 2 or more amino acid sequences are associated with Pfa2p (SEQ ID NO: 4 or sequence identification 50 201038734: 71) amino acid Contains 8 % identity, which contains one or more domains of PUFA synthetase, eg KS domain (SEQ ID NO: 3〇 or Sequence ID: 103), CLF domain (SEQ ID NO: 32 or sequence Identification number: 105), AT field (sequence identification number: 34 or sequence identification number: 107) 'ER field (sequence identification number: 36 or sequence identification number: 109), and combinations thereof, etc. In some specific examples, The polypeptide comprises one or more amino acid sequences in pfa2p (SEQ ID NO: 5, or Sequence ID: 71) comprising one or more domains of PUFA synthetase, including any individual domain A combination of one or more copies with one or more copies of any other individual field. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleocapamic acid sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence in the pfa3p (SEQ ID NO: 6 or Sequence ID: 73) At least 8% homologous amino acid sequence comprising - or more than one pUFA synthetase domain. For some specific shots, the polypeptide comprises at least 8% identity with the amino acid sequence (SEQ ID NO: 6 or SEQ ID NO: 73) in Pfa3p. t-amine oxime- or more PUFA synthase Fields, for example: DH field (eg sequence identification number: 38, sequence identification number: 4〇, sequence identification number: in, or sequence identification number: 113), ER field (sequence identification number: 42 or sequence identification number: ι15 ), and its group of eight. In some embodiments, the polypeptide comprises each of the at least two or more amino acid sequences of the two or more amino acid sequences, and pfa3p (SEQ ID NO: 6 or Sequence ID: The amino acid sequence in 73) has 8 % homology, which comprises - or more than one of the PUFA synthetase domains. In some specific examples 51 201038734

中，該至少2個或更多個胺基酸序列與Pfa3p (序列辨識編 號：6或序列辨識編號：73)中相同的胺基酸序列有80%的同 一性，其包含一或更多個PUFA合成酶領域。於一些具體例 中，該至少2個或更多個胺基酸序列與Pfa3p(序列辨識編 號：6或序列辨識編號：73)中不同的胺基酸序列有80%的同 一性，其各包含一或更多個PUFA合成酶領域。於一些具體 例中’該至少2個或更多個胺基酸序列與pfa3p (序列辨識編 號：6或序列辨識編號：73)中不同的胺基酸序列有80°/。的同 一性’其中當比較Pfa3p (序列辨識編號·· 6或序列辨識編 號：73)中的對應領域的等級時，該至少2個或更多個胺基 酸序列係位於該多肽中之相同的等級或不同的等級。於一 些具體例中’該至少2個或更多個胺基酸序列與Pfa3p (序列 辨識編號· 6或序列辨識編號：73)中的胺基酸序列有80%的 同一性，其包含—或更多個PUFA合成酶領域，例如：DH ：％、序列辨識編號：4◦、序列 辨識編*· 111 ’或序㈣識編號：U3)，ER領域(序列辨 識編5虎42或序列辨識編號：115)，以及其等之組合。於 匕具體例中，邊多肽包含Pfa3p(序列辨識編號：ό或序列 辨識編號：73)中沾, 個或更多個胺基酸序列，其包含一或 更多個PUFA合成醃 ^靖域，其包括任何個別的領域之一或更 多個副本與任何装 '、他個別的領域之一或更多個副本的組 合。 於一些具體例巾1 ’本發明係針對一核酸分子，其包含 編碼一多肽之多核钻 甘賤序列，其中該多肽包含與序列辨識 52 201038734 編號：2或序列辨識編號：69有至少80%的同一性之胺基酸 序列，以及其中該多肽包含選自於以下所構成的群組之 PUFA合成酶活性：KS活性、MAT活性、ACP活性、KR活 性、DH活性、以及其等之組合。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：8或序列辨識編號：75有至少80%的同一性之胺基酸 序列，以及其中該多肽包含KS活性。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：10或序列辨識編號：77有至少80%的同一性之胺基 酸序列，以及其中該多肽包含MAT活性。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：14、16、18、20、22、24、81、83、85、87、89、 91、93、95、97或99的任何一者有至少80%的同一性之胺 基酸序列，以及其中該多肽包含ACP活性。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：12或序列辨識編號：79有至少80%的同一性之胺基 酸序列，以及其中該多肽包含ACP活性。 於一些具體例中，本發明係針對核酸分子，其包含編 碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識編 號：12有至少80%的同一性之胺基酸序列，以及其中該多 53 201038734 肽包含ACP活性。於一些具體例中，該胺基酸序列與序列 辨識編號：12中的胺基酸序列有至少80%的同一性，其包 含1、2、3、4、5或6個ACP領域，其中該多肽包含與一或 更多個ACP領域關聯的ACP活性。序列辨識編號：14、16、 18、20、22與24為代表性胺基酸序列，其各包含序列辨識 編號：12中的單一 ACP領域。 於一些具體例中，本發明係針對核酸分子，其包含編 碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識編 號：79有至少80%的同一性之胺基酸序列，以及其中該多 肽包含ACP活性。於一些具體例中，該胺基酸序列係與序 列辨識編號：79中的胺基酸序列有至少80%的同一性，其 包含1、2、3、4、5、6、7、8、9或10個ACP領域，其中該 多肽包含與一或更多個ACP領域關聯的ACP活性。序列辨 識編號：81、83、85、87、89、91、93、95、97及99為代 表性胺基酸序列，其各包含序列辨識編號：79中的單一ACP 領域。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：26或序列辨識編號：101有至少80%的同一性之胺基 酸序列，以及其中該多肽包含KR活性。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：28或序列辨識編號：119有至少80%的同一性之胺基 酸序列，以及其中該多肽包含DH活性。 54 201038734 於一些具體例中，本發明係針對核酸分子，其包含編 碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識編 號：4或序列辨識編號：71有至少80%的同一性之胺基酸序 列，以及其中該多肽包含選自於以下所構成的群組之PUFA 合成酶活性：KS活性、CLF活性、AT活性、ER活性、以及 其等之組合。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：30或序列辨識編號：103有至少80%的同一性之胺基 酸序列，以及其中該多肽包含KS活性。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：32或序列辨識編號：105有至少80%的同一性之胺基 酸序列，以及其中該多肽包含CLF活性。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：34或序列辨識編號：107有至少80%的同一性之胺基 酸序列，以及其中該多肽包含AT活性。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：36或序列辨識編號：109有至少80%的同一性之胺基 酸序列，以及其中該多肽包含ER活性。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 55 201038734 編號：6或序列辨識編號：73有至少80%的同一性之胺基酸 序列，以及其中該多肽包含選自於以下所構成的群組之 PUFA合成酶活性：DH活性、ER活性、以及其等之組合。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：38有至少80%的同一性之胺基酸序列，以及其中該 多肽包含DH活性。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：40有至少80%的同一性之胺基酸序列，以及其中該 多肽包含DH活性。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：111有至少80%的同一性之胺基酸序列，以及其中該 多肽包含DH活性。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：113有至少80%的同一性之胺基酸序列，以及其中該 多肽包含DH活性。 於一些具體例中，本發明係針對一核酸分子，其包含 編碼一多肽之多核苷酸序列，其中該多肽包含與序列辨識 編號：42或序列辨識編號：115有至少80%的同一性之胺基 酸序列，以及其中該多肽包含ER活性。 於一些具體例中，該等核酸分子包含與本文中報導的 56 201038734 ❹Wherein the at least two or more amino acid sequences are 80% identical to the same amino acid sequence in Pfa3p (SEQ ID NO: 6 or Sequence ID: 73), which comprises one or more The field of PUFA synthetase. In some embodiments, the at least two or more amino acid sequences are 80% identical to the amino acid sequence different from Pfa3p (SEQ ID NO: 6 or Sequence ID: 73), each of which comprises One or more fields of PUFA synthetase. In some embodiments, the at least two or more amino acid sequences differ from the amino acid sequence of pfa3p (SEQ ID NO: 6 or Sequence ID: 73) by 80°. Identity of the 'where is the ratio of the corresponding domain in Pfa3p (SEQ ID NO: 6 or Sequence ID: 73), the at least 2 or more amino acid sequences are identical in the polypeptide Level or different level. In some embodiments, the at least two or more amino acid sequences are 80% identical to the amino acid sequence in Pfa3p (SEQ ID NO: 6 or Sequence ID: 73), which comprises - or More PUFA synthetase fields, for example: DH: %, sequence identification number: 4◦, sequence identification code *· 111 ' or sequence (four) identification number: U3), ER field (sequence identification code 5 tiger 42 or sequence identification number :115), and combinations of them. In a specific example, the side polypeptide comprises Pfa3p (SEQ ID NO: ό or SEQ ID NO: 73), and one or more amino acid sequences comprising one or more PUFA synthetic salts, It includes one or more copies of any individual field and any combination of one, one or more of its individual fields. In some specific embodiments, the invention relates to a nucleic acid molecule comprising a polynuclear diamond sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% with sequence identification 52 201038734 number: 2 or sequence identification number: 69. The amino acid sequence of identity, and wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of KS activity, MAT activity, ACP activity, KR activity, DH activity, and combinations thereof. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% identity to sequence identification number: 8 or sequence identification number: 75. An amino acid sequence, and wherein the polypeptide comprises KS activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% identity to sequence identification number: 10 or sequence identification number: 77. An amino acid sequence, and wherein the polypeptide comprises MAT activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises and the sequence identification number: 14, 16, 18, 20, 22, 24, 81, 83 Any one of 85, 87, 89, 91, 93, 95, 97 or 99 having at least 80% identity amino acid sequence, and wherein the polypeptide comprises ACP activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% identity to sequence identification number: 12 or sequence identification number: 79. An amino acid sequence, and wherein the polypeptide comprises ACP activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 12, and wherein The poly 53 201038734 peptide contains ACP activity. In some embodiments, the amino acid sequence is at least 80% identical to the amino acid sequence of Sequence Identification Number: 12, comprising 1, 2, 3, 4, 5 or 6 ACP domains, wherein The polypeptide comprises ACP activity associated with one or more ACP domains. Sequence Identification Numbers: 14, 16, 18, 20, 22, and 24 are representative amino acid sequences each comprising a single ACP domain in Sequence Identification Number: 12. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 79, and wherein The polypeptide comprises ACP activity. In some embodiments, the amino acid sequence is at least 80% identical to the amino acid sequence of Sequence Identification Number: 79, and comprises 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 ACP domains, wherein the polypeptide comprises ACP activity associated with one or more ACP domains. Sequence Identification Numbers: 81, 83, 85, 87, 89, 91, 93, 95, 97, and 99 are representative amino acid sequences each comprising a single ACP domain in Sequence Identification Number: 79. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% identity to sequence identification number: 26 or sequence identification number: 101. An amino acid sequence, and wherein the polypeptide comprises KR activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% identity to sequence identification number: 28 or sequence identification number: 119. An amino acid sequence, and wherein the polypeptide comprises DH activity. 54 201038734 In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% identity to sequence identification number: 4 or sequence identification number: 71 The amino acid sequence, and wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of KS activity, CLF activity, AT activity, ER activity, and combinations thereof. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% identity to sequence identification number: 30 or sequence identification number: 103. An amino acid sequence, and wherein the polypeptide comprises KS activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% identity to sequence identification number: 32 or sequence identification number: 105. An amino acid sequence, and wherein the polypeptide comprises CLF activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% identity to sequence identification number: 34 or sequence identification number: 107. An amino acid sequence, and wherein the polypeptide comprises AT activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% identity to sequence identification number: 36 or sequence identification number: 109. An amino acid sequence, and wherein the polypeptide comprises ER activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% identical to the sequence recognition 55 201038734 number: 6 or sequence identification number: 73 Amino acid sequence, and wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of DH activity, ER activity, and combinations thereof. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to Sequence ID: 38, and Wherein the polypeptide comprises DH activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence ID: 40, and Wherein the polypeptide comprises DH activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 111, and Wherein the polypeptide comprises DH activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 113, and Wherein the polypeptide comprises DH activity. In some embodiments, the invention is directed to a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 80% identity to sequence identification number: 42 or sequence identification number: 115. An amino acid sequence, and wherein the polypeptide comprises ER activity. In some embodiments, the nucleic acid molecules comprise as described herein 56 201038734 ❹

該多核苷酸序列有至少大約80%、85%’或90%同一性之多 核苷酸序列，或包含與本文中報導的該多核苷酸序列有至 少大約 95%、96%、97%、98%、99%，或 100% 同一性之多 核苷酸序列。如熟悉此藝者知道的，術語"百分比同一性" 為2或更多個胺基酸序列或2或更多個多核苷酸序列之間的 關係，如同藉由比較該等序列決定的。於本技藝中，’’同一 性"亦意指介於胺基酸或多核苷酸序列之間的序列關係的 程度，例如本情況可為，係如同藉由此等序列串之間的配 對來決定。 關於一核酸分子，其具有’舉例而言’與本發明的一 參考多核苷酸序列有至少95%的"同一性(identical)"之多核 苷酸序列，其係意指該核酸分子的多核苷酸序列係與該參 考序列為完全相同的’除了在參考多核苷酸序列的每各1〇〇 個核苷酸中該多核苷酸序列能包括多至5個核苷酸差異。換 言之，要得到一核酸分子’其具有與參考多核苷酸序列有 至少95%的同一性之多核苷酸序列，該參考序列中多至5% 的核苷酸可以被刪失或是用另一個核苷酸取代的’或是該 參考序列内的一些核苷酸（多至5%的總核苷酸)能被插入至 該參考序列之内。 就實務而言，任何特定的多核苷酸序列或胺基酸序列 是否與本發明的多核苷酸序列或胺基酸序列有至少80°/〇、 85%、90%、95%、96%、97%、98%或99%同一性能慣常地 使用已知的電腦程式予以決定。一種用於決定介於一詢問 序列（本發明的序列）以及一主題序列之間的最佳全面的匹 57 201038734 配的方法能使用序列的排列以及計算同一性分數來決定。 排列係使用電腦程式AlignX來進行，其係來自Invitr0gen (www.invitrogen.com)的載體NTI Suite 10.0套裝軟體之組 份。排列係使用ClustalW排列（Thompson, J.D.等人，Nucl. Acids Res. 22: 4673-4680 (1994))來執行胺基酸與多核苷酸 序列排列二者。各別使用系統預設計分矩陣m〇s腿62mt2 和swgapdnamt用於胺基酸與多核苷酸序列排列。關於胺基 酸序列，系統預設空格開放罰分為10以及空格擴展罰分為 0.1。關於多核苷酸序列’系統預設空格開放罰分為15以及 空格擴展罰分為6.66。 本發明係針對一種經單離的核酸分子，其包含編碼一 多肽之多核苷酸序列’該多肽包含選自於以下所構成的群 組之PUFA合成酶活性：KS活性、MAT活性、ACP活性、 KR活性、CLF活性、AT活性、ER活性、DH活性、以及其 等之組合’其中該多核苷酸係於嚴苛條件下與以上說明的 該等多核苷酸序列的任一者之該互補物雜交。 當該核酸分子的一單股形式能於溫度和溶液的離子強 度之合適的條件下黏合至另一核酸分子時，一核酸分子為” 可雜交π至另一個核酸分子，例如：cDNA、基因體DNA， 或是RNA的。雜交與清洗條件係熟知且舉例說明的。參見， 例如，Sambrook J. and Russell D. 2001. Molecular cloning: A laboratory manual，3rd edition。Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York。溫度與離 子強度條件決定雜交的n嚴苛性"。可以調整嚴苛條件以篩 58 201038734 選出中度相似的片段，例如：來自略微相關的有機體之同 源性序列，至高度相似的片段，例如：來自緊密相關的有 機體之複製功能性酵素基因。雜交後清洗的會決定嚴苛條 件。一組條件使用一系列的清洗，開始用6χThe polynucleotide sequence has a polynucleotide sequence of at least about 80%, 85% ' or 90% identity, or comprises at least about 95%, 96%, 97%, 98 of the polynucleotide sequence reported herein. A polynucleotide sequence of %, 99%, or 100% identity. As is familiar to those skilled in the art, the term "percent identity" is the relationship between two or more amino acid sequences or two or more polynucleotide sequences, as determined by comparing such sequences. . In the present technique, ''identity" also means the degree of sequence relationship between amino acid or polynucleotide sequences, for example, in this case, as if by pairing between such sequence strings To decide. With respect to a nucleic acid molecule, which has, by way of example, a polynucleotide sequence of at least 95% "identical" with a reference polynucleotide sequence of the invention, which means The polynucleotide sequence is identical to the reference sequence 'except for each nucleotide sequence of the reference polynucleotide sequence, the polynucleotide sequence can include up to 5 nucleotide differences. In other words, a nucleic acid molecule is obtained which has a polynucleotide sequence which is at least 95% identical to the reference polynucleotide sequence, and up to 5% of the nucleotides in the reference sequence can be deleted or used in another Nucleotide-substituted 'or some of the nucleotides within the reference sequence (up to 5% of the total nucleotides) can be inserted into the reference sequence. In practice, whether any particular polynucleotide sequence or amino acid sequence is at least 80°/〇, 85%, 90%, 95%, 96% with the polynucleotide sequence or amino acid sequence of the invention, 97%, 98%, or 99% of the same performance is routinely determined using known computer programs. One method for determining the best overall fit between an interrogation sequence (the sequence of the invention) and a subject sequence can be determined using the alignment of the sequences and the calculation of the identity score. The alignment was performed using the computer program AlignX, which is a component of the carrier NTI Suite 10.0 software package from Invitr0gen (www.invitrogen.com). The alignment was performed using the ClustalW arrangement (Thompson, J. D. et al., Nucl. Acids Res. 22: 4673-4680 (1994)) to perform both amino acid and polynucleotide sequence alignment. The system pre-designed sub-matrix m〇s legs 62mt2 and swgapdnamt are used for the amino acid and polynucleotide sequence alignment, respectively. Regarding the amino acid sequence, the system presets a space open penalty of 10 and a space extension penalty of 0.1. Regarding the polynucleotide sequence, the system presets a space open penalty of 15 and a space extension penalty of 6.66. The present invention is directed to an isolated nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide comprising a PUFA synthase activity selected from the group consisting of KS activity, MAT activity, ACP activity. , KR activity, CLF activity, AT activity, ER activity, DH activity, and combinations thereof, wherein the polynucleotide is complementary to any of the polynucleotide sequences described above under severe conditions Hybridization. When a single-stranded form of the nucleic acid molecule can be bonded to another nucleic acid molecule under suitable conditions of temperature and ionic strength of the solution, a nucleic acid molecule can be hybridized to another nucleic acid molecule, such as cDNA, gene body. DNA, or RNA. Hybridization and washing conditions are well known and exemplified. See, for example, Sambrook J. and Russell D. 2001. Molecular cloning: A laboratory manual, 3rd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor , New York. Temperature and ionic strength conditions determine the n-rigidity of hybridization. The harsh conditions can be adjusted to screen 58 201038734 to select moderately similar fragments, eg, homologous sequences from slightly related organisms, to a high degree of similarity Fragments, such as: replication of functional enzyme genes from closely related organisms. Cleaning after hybridization determines harsh conditions. A set of conditions uses a series of cleanings, starting with 6χ

SSCO.5% SDS 在室溫下歷時15 min，接而用2X ssc ' 〇 5% SDS在45〇c下 重複歷時30 min，以及接而用〇 2X ssc、〇 5% 3]〇8在5〇0(^ 下重複二次歷時3〇 minD至於更嚴苛的條件，在較高的溫 0 錄行清洗，其巾清洗係與以上完全相同，除了最後二次 30 min於予以〇·2χ ssc、〇5%麗中之清洗的溫度增加至 C之外另.組尚度嚴苛的條件係使用在65°C下於〇.IX SSC’ 0.1/〇 SE)S中之二次最後的清洗。額外一組的高度嚴苛 . 的條件係定義為：在65°Cf於0.1X SSC、0.1% SDS中雜交 以及用 2X SSC、〇.l% SDS接著〇 ιχ ssc、〇 1% sds予以清 洗。 本發明係針對-種經單離的核酸分子，其包含與以上 〇 °兒明的4等多核聽相的任—者完全互補的多核紐序 列。術語"互補的"係料說·此雜交的料酸驗基 之間的關係。舉例而言，關於DNA，腺苦係互補於胸腺„密 啶以及胞嘧啶係互補於鳥糞嘌呤。 於某些具體例中，該多核苦酸或核酸為DNA。於DNA 的情況I，-核酸分子，其包含編碼一多肽之多核苦酸序 苇月匕括與一或更多個編碼區域操作上關聯之啟動 子及/或其他的轉錄或是轉譯控制元素。操作上_性為當 一基因產物(例如，—多肽)的編碼區域係與-或更多個關聯 59 201038734 的調控的序列時，以此一方式使得該基因產物的表現係調 控序列的影響下或是調控序列的控制下。二個DNA片段(例 如一多肽編碼區域以及與之關聯的一啟動子）為"操作上關 聯的"，設若啟動子功能的誘導導致編碼所欲的基因產物之 mRNA的轉錄以及設若介於二個DNA片段之間的鍵聯本質 不干擾表現調控的序列指示基因產物表現的能力或是干擾 待轉錄之DNA模板的能力。因而，—啟動子區域會與編碼 -多肽之多核苦酸序列操作上關聯，設若該啟動子能夠造 成那個多核苷酸序列的轉錄。該啟動子可以是一細胞專一 性啟動子，其僅於預定的細胞内指*DNA之實質的轉錄。 一般而言，一編碼區域係座落於一啟動子的3，。啟動子可以 其等之全體係衍生自天然的基因，或是由自然界發現之不 同的啟動子衍生的不同的元素所組成的，或者甚至包含合 成性DNA區段。熟悉此藝者所了解的，不同的啟動子能指 不不同的組織或細胞類型内，或在不同的發展階段的基因 之表現，或是對不同的環境或生理條件反應的基因之表 現。啟動子，其造成一基因在大部分時間、於多數細胞類 型内表現，通常被稱為"構成性啟動子"。進一步承認因於 多數情況下調控的序列之確實的邊界並未完全地界定，不 同長度的DNA片段能具有同一的啟動子活性。一啟動子一 般以轉錄起始位置旁的其之3,端為界，以及向上游(5，方向） 延伸以包括起始背景資料以上之可偵測的位準之轉錄所必 須的最小數目的鹼基或元素。於啟動子内會發現_轉錄起 始位置（合宜地界定，舉例而言，用核酸酶S1予以繪圖），以 60 201038734 及負責RNA聚合酶的結合之蛋白結合領域（一致序列）。 適當的調控的區域包括核酸區域係座落於一編碼區域 的上游(5'非編碼序列）、一編碼區域内，或是一編碼區域下 游(3'非編碼序列），以及其影響關聯的編碼區域之轉錄、 RNA加工或是安定性，或是轉譯。調控的區域能包括：啟 動子、轉譯引導序列、RNA加工位址、效應子結合位置， 以及莖-環結構。除一啟動子以外，其他的轉錄控制元素， _ 舉例而言：增強子、操作子、抑制子，以及轉錄終止信號， 〇 可以與該多核苷酸操作上關聯以指示細胞專一性轉錄。該 編碼區域的邊界係精由5'(胺基）端處之起始密碼子及3'(魏 基)端處之轉譯終止密碼子來決定。一編碼區域能包括，但 不限於：原核區域、來自mRNA的cDNA、基因體DNA分子、 合成性DNA分子，或是RNA分子。設若該編碼區域係預期 於一真核細胞内之表現，一聚腺苷酸化的訊號及轉錄終止 序列通常會座落於該編碼區域的3·。 q 於本發明的某些態樣中，具有至少20個鹼基、至少30 個鹼基，或是至少50個鹼基以及雜交至本發明的多核苷酸 序列之多核苷酸序列可以使用作為PCR引子。典型地，於 PCR型擴增的技術中，該等引子具有不同的序列以及為彼 此互補的。取決於所欲的測試條件，該等引子的序列應該 被設計以提供標的核酸之有效且忠實的複製。PCR引子設 計的方法為常見的以及係本技藝所熟知的。一般，本序列 之二個短的區段能使用於聚合酶連鎖反應（PCR)流程之中 以擴增編碼來自DNA或RNA的同源性基因之更長的核酸片 61 201038734 段。聚合酶連鎖反應也可以執行於經選殖的核酸片段之 庫，其中一引子的序列係自本核酸片段所衍生’以及另— 個的引子的序列利用聚腺苷酸束的存在而至編碼微生物基 因的mRNA前驅物之3’末端。任擇地，第二引子的序列可以 根據該選殖載體所衍生的序列。 此外，專一性引子可以被設計且用來擴增本序列之部 分或全長。形成的擴增產物可以在擴增反應期間直接地予 以標記或是在擴增反應之後予以標記，以及被使用作為探 針以於合適的嚴苛條件下單離全長DNA片段。 因而’本發明的核酸分子能使用來單離基因，該等基 因係編碼來自相同或其他的物種或細菌的物種之同源性蛋 白質。使用序列依賴性流程之同源性基因的單離係本技藝 所熟知的。序列依賴性流程的實例包括，但不限於：核酸 的雜交的方法’以及DNA和RNA擴增的方法，如藉由各種 各樣的使用核酸擴增的技術所舉例說明的（例如：聚合酶連 鎖反應，Mullis等人，美國專利案第4,683,202號；連接酶連 鎖反應(LCR) (Tabor, S.等人，Proc. Acad· Sci· USA 82: 1074 (1985));或股置换擴增（SDA ; Walker等人，Proc. Natl. Acad. Sci. U.S.A. 89: 392 (1992))〇 於一些具體例中，本發明之經單離的核酸分子係用來 單離來自其他有機體之同源的核酸分子俾以鑑定產生相似 的或是改良的PUFA形態之PUFA合成酶。於一些具體例 中，本發明之經單離的核酸分子係用來單離來自其他有機 體之同源的核酸分子，其等係涉及生產高量的DHA。 62 201038734 本發明的核酸分子亦包含多核苷酸序列，其編碼PUFA 合成酶基因、PUFA合成酶基因的領域，或是被符合框架地 融合至一標誌序列之PUFA合成酶基因的片段，該標誌序列 允許本發明的多肽之偵測。標誌序列包括熟悉此藝者所知 道之營養缺陷型的或是顯性的標誌，例如：ZEO (吉歐黴素 (Zeocin))、NEO (G418)、潮黴素(hygromycin)、石申華、HPH、 NAT，以及類似物。 本發明亦包含該PUFA合成酶基因的變異體。變異體可 含有編碼區域、非編碼區域，或是二者内的改變。多核苷 酸序列的變異體之實例含有產生緘默取代、添加，或是删 失之改變，但是不改變編碼的多肽之性質或活性。於某些 具體例中，多核苷酸序列變異體係藉由由於遺傳密碼的簡 併之緘默取代來生產。於進一步的具體例中，多核苷酸序 列變異體可以為了各種各樣的原因而生產，例如，為了一 特定的宿主之最佳化密碼子的表現(例如，改變該破囊壺菌 m RN A内的密碼子成為其他有機體(例如大腸桿菌或是啤酒 酵母菌少ces cerev/s/ae))所偏好的該等）。 本發明中亦提供對偶基因變異體、直向同源物，及/或 種同源物。能使用本技藝所知道的程序、使用本文中揭示 的序列之資訊來得到本文中說明的基因之全長的基因、對 偶基因變異體、剪切變異體、全長的編碼部分、直向同源 物，及/或種同源物。舉例而言，對偶基因變異體及/或種同 源物可以藉由製作來自本文中提供的序列之適當的探針或 引子而予以單離及鑑定，以及以篩選用於對偶基因變異體 63 201038734 及/或所欲的同源物之適當的核酸來源。 本發明係針對一種重組型核酸分子，其包含：以上說 明的該等核酸分子的任一者或其等之組合以及一轉錄控制 序列。於一些具體例中，該重組型核酸分子為一重組塑載 體。 本發明係針對一種製造一重組型載體的方法’其包 含：將如本文中說明的一或更多個經單離的核酸分子插入 至一載體中。 本發明的載體，舉例而言’能是一選殖載體或一表現 載體。該載體可以是，舉例而言，以一質體、一病毒顆粒、 一噬菌體等等的形式。 本發明的多核苷酸序列能被包括於用於表現一多肽之 各種各樣的表現載體之任何一者内。此等載體包括染色 體、非染色體，以及合成性DNA或是RNA序列，例如，SV40 的衍生物；細菌質體；以及酵母菌質體。然而，能使用熟 悉此藝者所知道之任何其他合適的載體。 合適的DNA序列能藉由各種各樣的程序被插入至載體 之内。一般而言，該DNA序列係藉由本技藝所知 被插入至合適的限制性内生核酸酶，序 以及其餘的被視為落在熟悉此藝者的_内。此等的程序 本發明亦包括重組型建構物，其包含以上說 更多個多核苷酸序列 该等建構物包含一载 明的 體，例如 或 或是反向_插二:::更^ 態樣 64 201038734 中，該建構物進-步包含操作上關聯至該序列之調控的序 列’包括，舉例而言，—啟動子。大數目的適當載體及啟 動子係也悉此藝者知道的，以及為商業上可得的。 多肽 本發明係針對經單離的多肽，其包含PUFA合成酶蛋白 與領域之胺基酸序列，其等係自以ATCC寄存編號ρτΑ_9695 及ΡΤΑ-10212所寄存之經單離的微生物所衍生的。 0 當使用於本文中，術語”多肽"係預期要包含一單一的，， 夕肽及複數的多肽”，以及係提及由經由醯胺鍵（亦知道為 肽鍵）而成直線地連結的單體(胺基酸)所組成的一分子。術 - S吾多肽'•係提及2或更多個胺基酸之任何的鏈(chain)威鍊 (chains)，以及非為提及一特定長度的產物。因而，用來槔 及2或更多個胺基酸的鏈(chain)或鏈(也以⑽)之肽、二肽、彡 肽、寡肽、"蛋白質”、”胺基酸鏈”或是任何其他的術語係包 括於"多肽”的定義内，以及術語"多肽"能使用來代替此等樹 Q 語的任何一者或是與此等術語的任何一者互相交換使用。 術語"多肽"亦預期要提及該多肽的表現後修飾之產物，私 括，沒有限制：醣化、乙醯化、磷酸化、醯胺化、藉由已 知的保護基/封阻基之衍生化' 蛋白分解分裂，或是藉由井 天然存在的胺基酸之修飾。 如本文中說明的多肽可包括其等之片段、變異體，成 是衍生分子，沒有限制。術語"片段變異體"、"衍生物" 以及類似物"當參照一多肽時係包括保留至少一些生物的 活性之任何多肽。多肽片段能包括：蛋白分解片段、冊J夹 65 201038734 片段，以及當被遞送至一動物時更容易達到作用位址的片 段。多肽片段進一步包括該多肽的任何部分，其包含天然 的多肽之抗原性或是免疫性抗原決定位，包括直線，以及 三維的抗原決定位。多肽片段能包含變異體區域，其包括 如以上說明的片段，以及也包含具有由於胺基酸取代、刪 失，或是插入而改變的胺基酸序列之多肽。變異體能天然 地發生，例如一對偶基因變異體。按”對偶基因變異體”係 預期基因之任擇的形式，其係佔據一有機體的染色體之給 定的位址。非天然存在的變異體可以使用本技藝已知的致 突變技術予以生產。本發明的多肽片段能包含守恒性或非 守恆性胺基酸取代、刪失，或是添加。變異體多肽於本文 中亦能稱為”多肽類似物”。本發明的多肽片段也能包括衍 生分子。當使用於本文中，一多肽或一多肽片段的’’衍生物 "係提及一主題多肽，其具有藉由功能性側基的反應所化學 上衍生的一或更多個殘基。”衍生物"也包括該等肽，其等 含有20個標準的胺基酸之一或更多個天然存在的胺基酸衍 生物。舉例而言，4-羥脯胺酸可以取代脯胺酸；5-羥離胺酸 可以取代離胺酸；3-曱基組胺酸可以取代組胺酸；類絲胺 酸可以取代絲胺酸；以及鳥胺酸可以取代離胺酸。 本發明的多肽能藉由本發明的該等核酸分子的任一者 所編碼。 本發明係針對經單離的多肽，其包含與Pfalp (序列辨 識編號：2或序列辨識編號：69)、Pfa2p (序列辨識編號：4 或序列辨識編號：71)、Pfa3p(序列辨識編號：6或序列辨識 66 201038734 編號：73)，以及其等之組合的胺基酸序列有至少8〇。/。的同 一性之胺基酸序列，其中該等多肽包含一或更多PUFA合成 酶活性。 本發明係針對多肽，其包含與本發明的PUFA合成酶之 一或更多個PUFA合成酶領域的胺基酸序列有至少8〇%的同 一性之胺基酸序列。 於一些具體例中，本發明係針對多肽，其包含與pfal 0 (序列辨識編號：2或序列辨識編號：69)中的胺基酸序列有 至少80%的同一性之胺基酸序列，包含一或更多個puFA合 成_領域。於一些具體例中，該多肽包含與Pfal (序列辨識 . 編號· 2或序列辨識編號：69)中的胺基酸序列有至少80%的 同一性之胺基酸序列，包含一或更多個puFA合成酶領域， 例如：ks領域(序列辨識編號：8或序列辨識編號：75)， MAT領域(序列辨識編號：1〇或序列辨識編號：77)，ACp 領域(例如.序列辨識編號：14、16、18、20、22、24、81、 Q 83、85、87、89、91、93、95、97或99的任何一者），2個 或更夕個ACP領域的組合，例如：2、3、4、5、6、7、8、 9或10個ACP領域，包括串聯領域(序列辨識編號：12或序列 辨識編號：79，以及其等之部分），KR領域(序列辨識編號： 26或序列辨識編號：101)，DH領域(序列辨識編號：28或序 列辨識編號：119)，以及其等之組合。於一些具體例中， 該多肽包含2個或更多個胺基酸序列，其中該至少2個或更 多個胺基酸序列的各個係與Pfalp (序列辨識編號：2或序列 辨識編號：69)中的胺基酸序列有80%的同一性，包含一或 67 201038734 更多個PUFA合成酶領域。於一些具體例中’該至少2個或 更多個胺基酸序列與Pfalp (序列辨識編號：2或序列辨識編 號：69)中相同的胺基酸序列有80%的同一性’包含一或更 多個PUFA合成酶領域。於一些具體例中，該至少2個或更 多個胺基酸序列與Pfalp (序列辨識編號：2或序列辨識編 號：69)中不同的胺基酸序列有80%的同一性，其各包含_ 或更多個PUFA合成酶領域。於一些具體例中，該至少2個 或更多個胺基酸序列與Pfalp (序列辨識編號：2或序列辨識 編號：69)中不同的胺基酸序列有80%的同一性，其中當比 較Pfalp(序列辨識編號：2或序列辨識編號：69)中的對應領 域的等級時’該至少2個或更多個胺基酸序列係位於該多肽 中之相同的等級或不同的等級。於一些具體例中，該至少2 個或更多個胺基酸序列與Pfalp(序列辨識編號：2或序列辨 識編號：69)中的胺基酸序列有80%的同一性，包含一或更 多個PUFA合成酶領域’例如：Ks領域(序列辨識編號：8 或序列辨識編號：75)，MAT領域(序列辨識編號：1〇或序列 辨識編號：77) ’ ACP領域(例如：序列辨識編號：14、16、 18、20'22、24、8卜83、85、87、89、91、93、95、97 或99的任何—者），2、3、4、5、6、78、9或1()個心領 域的組合，包括串聯領域(序列辨識編號：12或序列辨識編 號：79，以及其等之部分），KR領域(序列辨識編號^或 序列辨識編號]G1)，DH領域(序列辨識編號：辦序列辨 識編號以及其等之組合。於—些具體例中，該多 肽包含PfaiP(序列辨識編號：2或序列辨識編號：69)中的丄 68 201038734 個或更多個胺基酸序列’包含-或更多個PUFA合成酶領 域其包括任何個別的領域之一或更多個副本與任何其他 個別的領域之一或更多個副本的組合。 於一些具體例中，本發明係針對多肽，其包含與pfa2p (序列辨識、《 : 4或相觸職：川巾的絲酸序列有 至>、80/。的同—性之胺基酸序列，包含—或更多個puFA合 成酶領域。於—些具體例中，該纽包含與Pfa2p中的胺基 酉文序列（序列辨識編號：4或序列辨識編號：71)有至少 的同一性之胺基酸序列，包含一或更多個puFA合成酶領 域，例如：KS領域（序列辨識編號：3〇或序列辨識編號： 103)，CLF領域(序列辨識編號：32或序列辨識編號：1〇5)， AT領域(序列辨識編號：34或序列辨識編號：1〇7)，ER領域 (序列辨識編號：36或序列辨識編號：1〇9)，以及其等之組 合。於一些具體例中，該多肽包含2個或更多個胺基酸序 列’其中該至少2個或更多個胺基酸序列的各個係與pfa2p (序列辨識編號：4或序列辨識編號：71)中的胺基酸序列有 80%的同一性，包含一或更多個PUFA合成酶領域。於一些 具體例中’該至少2個或更多個胺基酸序列與Pfa2p (序列辨 識編號：4或序列辨識編號：71)中相同的胺基酸序列有80〇/〇 的同一性。於一些具體例中，該至少2個或更多個胺基酸序 列與Pfa2p(序列辨識編號：4或序列辨識編號：71)中不同的 胺基酸序列有80%的同一性，其各包含一或更多個PUFA合 成酶領域。於一些具體例中，該至少2個或更多個胺基酸序 列與Pfa2p(序列辨識編號：4或序列辨識編號：71)中不同的 69 201038734 胺基酸序列有80〇/。的同一性，其中當比較pfa2p(序列辨識編 唬.4或序列辨識編號：71)中的對應領域的等級時，該至 J 2個或更多個胺基酸序列係位於該多肽中之相同的等級 或不同的等級。於一些具體例中，該至少2個或更多個胺基 酸序列與Pfa2p(序列辨識編號：4或序列辨識編號 :71)中的 胺基酸序列有80%的同一性，包含一或更多個puFA合成酶 領域，例如：KS領域(序列辨識編號：3〇或序列辨識編號： 1〇3)，CLF領域(序列辨識編號：32或序列辨識編號h〇5)， AT領域(序列辨識編號：34或序列辨識編號：1〇7)，er領域 (序列辨識編號：36或序列辨識編號：1〇9)，以及其等之組 合。於一些具體例中，該多肽包含Pfa2p(序列辨識編號：4 或序列辨識編號：71)中的1個或更多個胺基酸序列，包含 一或更多個PUFA合成酶領域，其包括任何個別的領域之一 或更多個副本與任何其他個別的領域之一或更多個副本的 組合。 於些具體例中，本發明係針對多肽，其包含與Pfa3p (序列辨識編號：6或序列辨識編號：73)中的胺基酸序列有 至少80%的同一性之胺基酸序列，包含一或更多個puFA合 成酶領域。於一些具體例中，該多肽包含與pfa3p(序列辨識 編號：6或序列辨識編號：73)中的胺基酸序列有至少8〇%的 同-性之胺基酸序列，包含-或更多個PUFA合成酶領域， 例如：DH領域(例如：序列辨識編號：38、序列辨識編號： 40、序列辨識編號：111 ’或序列辨識編號u13)，er領域 (序列辨識編號：42或序列辨識編號：115)，以及其等之組 70 201038734 合。於一些具體例中’該多肽包含2個或更多個胺基酸序 列，其中該至少2個或更多個胺基酸序列的各個係與Pfa3p (序列辨識編號：6或序列辨識編號：73)中的胺基酸序列有 80%的同一性，包含一或更多個PUFA合成酶領域。於一些 具體例中’該至少2個或更多個胺基酸序列與pfa3p (序列辨 識編號：6或序列辨識編號：73)中相同的胺基酸序列有80% 的同一性，包含一或更多個PUFA合成酶領域。於一些具體 例中，該至少2個或更多個胺基酸序列與Pfa3p (序列辨識編 號：6或序列辨識編號：73)中不同的胺基酸序列有80%的同 一性，其各包含一或更多個PUFA合成酶領域。於一些具體 例中’該至少2個或更多個胺基酸序列與pfa3p (序列辨識編 號：ό或序列辨識編號：73)中不同的胺基酸序列有8〇%的同 一性’其中當比較pfa3P(序列辨識編號：6或序列辨識編號： 73)中的對應領域的等級時，該至少2個或更多個胺基酸序 列係位於該多肽中之相同的等級或不同的等級。於一些具 體例中，該至少2個或更多個胺基酸序列與Pfa3p (序列辨識 編號：6或序列辨識編號：73)中的胺基酸序列有80%的同一 性，包含—或更多個pUFA合成酶領域，例如：DH領域(例 如.序列辨識編號：38 '序列辨識編號：40、序列辨識編 號.ill，或序列辨識編號：113)，ER領域(序列辨識編號： 42或序列辨識編號：115)，以及其等之組合。於一些具體 例中，邊多肽包含Pfa3p(序列辨識編號：6或序列辨識編號： 73)中的1個或更多個胺基酸序列，包含一或更多個PUFA合 成_7員域，其包括任何個別的領域之一或更多個副本與任 71 201038734 何其他個別的領域之一或更多個副本的組合。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號2 :或序列辨識編號：69有至少80%的同一性之 胺基酸序列，其中該多肽包含選自於以下所構成的群組之 PUFA合成酶活性：KS活性、MAT活性、ACP活性、KR活 性、DH活性、以及其等之組合。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：8或序列辨識編號：75有至少80%的同一性之 胺基酸序列，其中該多肽包含KS活性。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：10或序列辨識編號：77有至少80%的同一性 之胺基酸序列，其中該多肽包含MAT活性。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：14、16、18、20、22、24、81、83、85、87、 89、91、93、95、97或99的任何一者有至少80%的同一性 之胺基酸序列，其中該多肽包含ACP活性。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：12或序列辨識編號：79有至少80%的同一性 之胺基酸序列，其中該多肽包含ACP活性。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：12内的胺基酸序列有至少80%的同一性之胺 基酸序列，其中該多肽包含ACP活性。於一些具體例中， 該胺基酸序列係與序列辨識編號：12中的胺基酸序列有至 少80%的同一性，其編碼1、2、3、4、5或6個ACP領域，其 72 201038734 中該多肽包含與一或更多個ACP領域關聯的acp活性。序 列辨識編號：14、16、18、20、22與24為代表性胺基酸序 列，其包含：序列辨識編號：12中的單一ACP領域。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：79内的胺基酸序列有至少80%的同一性之胺 基酸序列，其中該多肽包含ACP活性。於一些具體例中， 該胺基酸序列係與序列辨識編號：79中的胺基酸序列有至 ❹ 少80%的同一性，其包含1、2、3、4、5、6、7、8、9或1〇 個ACP領域，其中該多肽包含與一或更多個ACP領域關聯 的ACP活性。序列辨識編號：81、83、85、87、89、91、 93、95、97及99為代表性胺基酸序列，其包含：序列辨識 ' 編號：79中的單一 ACP領域。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號.26或序列辨識編號：1〇丨有至少8〇%的同一性 之胺基酸序列，其中該多肽包含KR活性。 〇 ☆一些具體例中，本發明係針對-多肽，其包含與序 列辨識編號：28或序列辨識編號：119有至少8〇%的同—性 之胺基酸序列，其中該多肽包含Dh活性。 於一些具體例中，本發明係針對-多肽，其包含與序 列辨識編號：4或序列辨識編號：71有至少議的同—性之 胺基酸序歹^其中該多肽包含選自於以下所構成的群組之 PUFA合成酶A I邮纟性、CLF^、at活性、er活性、 以及其等之組合。 於一些具體例中，本發明係斜斜一多狀，其包含與序 73 201038734 列辨識編號：30或序列辨識編號：103有至少80%的同一性 之胺基酸序列，其中該多肽包含KS活性。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：32或序列辨識編號：105有至少80%的同一性 之胺基酸序列，其中該多肽包含CLF活性。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：34或序列辨識編號：107有至少80%的同一性 之胺基酸序列，其中該多肽包含AT活性。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：36或序列辨識編號：109有至少80%的同一性 之胺基酸序列，其中該多肽包含ER活性。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：6或序列辨識編號：73有至少80%的同一性之 胺基酸序列，其中該多肽包含選自於以下所構成的群組之 PUFA合成酶活性：DH活性、ER活性、以及其等之組合。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：38有至少80%的同一性之胺基酸序列，其中 該多肽包含DH活性。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：40有至少80%的同一性之胺基酸序列，其中 該多肽包含DH活性。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：111有至少80%的同一性之胺基酸序列，其中 該多肽包含DH活性。 74 201038734 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：113有至少80%的同一性之胺基酸序列，其中 該多肽包含DH活性。 於一些具體例中，本發明係針對一多肽，其包含與序 列辨識編號：42或序列辨識編號：115有至少80%的同一性 之胺基酸序列，其中該多肽包含ER活性。 於一些具體例中，該等多肽包含與本文中報導的該胺 基酸序列有至少大約80%、85%，或90%同一性之胺基酸序 列，或包含與本文中報導的該胺基酸序列有至少大約 95%、96%、97%、98%、99%，或 100%同一性之胺基酸序 列。 關於一多肽’其具有，舉例而言，與本發明的一詢問 的胺基酸序列有至少95°/。的"同一性(identical)，，之胺基酸序 列’其係意指該主題多肽的胺基酸序列係與該詢問的胺基 酸序列為完全相同的，除了在詢問的胺基酸序列的每各1〇〇 個胺基酸中該主題多肽序列能包括多至5個胺基酸差異。換 言之’要得到一多狀，其具有與詢問的胺基酸序列有至少 95%的同-性之胺基酸序列，該主題多肽中多至5%的胺基 酸殘基可以被插入、冊彳失(indds)或是用另一個胺基酸取 代。该參考序列的此等改變能發生在該參考胺基酸序列的 胺基或羧基端的位置或是於該等終端位置之間的任何地 方’個別地散佈於該參考序狀巾的殘基歧散佈於該參 考序列的一或更多個的相連基團之中。 就實務而言，是否任何特定的多肽具有與本發明的胺 75 201038734 基酸序列有至少 80%、85%、90%、95%、96%、97%、98% 或99%同一性之胺基酸序列，譬如，能慣常地使用如上討 論的已知的電腦程式予以決定。如上討論的，一種用於決 定介於一詢問序列（本發明的序列）以及一主題序列之間的 最佳全面的匹配的方法能使用序列的排列以及計算同一性 分數來決定。排列係使用電腦程式AlignX來進行，其係來 自 Invitrogen(www.invitrogen.com)的載體NTI Suite 10.0套 裝軟體之組份。排列係使用ClustalW排列（J. Thompson等 人，Nucleic Acids Res. 22(22):4673-4680 (1994)來執行。使 用系統預設計分矩陣Blosum62mt2。系統預設空格開放罰分 為10以及空格擴展罰分為0.1。 於本發明之進一步的態樣中，核酸分子，其等與本文 揭示的多核苷酸序列有至少80%、85%、90%、95%、96%、 97%、98%或99%同一性之多核苷酸序列，編碼具有一或更 多PUFA合成酶活性之一多肽。具有一或更多PUFA合成酶 活性多肽展現出一或更多個的活性，其等相似於本發明的 PUFA合成酶之一或更多個的活性，但是不必然與本發明的 PUFA合成酶之一或更多個的活性完全相同。 當然，由於遺傳密碼的簡併，熟悉此藝者會立即承認 大部分的核酸分子，其等具有與本文中說明的多核苷酸序 列有至少 80〇/°、85°/。、90。/〇、95%、96%、97%、98%或99% 同一性之多核苷酸序列，會編碼"具有PUFA合成酶功能性 活性"的多肽。事實上’因此等多核苷酸序列的任何一者之 簡併性變異體全部編碼相同的多肽，於許多例子中，其能 76 201038734 藉由熟悉此藝者根據守恆性取代，以及守恆性功能性領 域，其多肽會展現出活性，之知識予以預測。於本發明的 某些態樣中，本發明的多肽與多核苷酸係以經單離的形式 提供，例如，經純化至均質性。任擇地，本發明的多肽與 多核苷酸可以藉由慣用的合成器予以合成地生產。 如本技藝所知道的介於二個多肽之間的"相似性"係藉 由比較該胺基酸序列和一第二多肽的序列之那裡的多肽的 守恆性胺基酸取代基予以決定。 Ο 於一些具體例中，本發明的多肽為一融合多肽。 當使用於本文中，π融合多肽’'意指一多肽，其包含經 由肽鍵成直線地連接至一第二多肽的一第一多肽。該第一 ' 多肽與該第二多肽可以是同一的或是不同的，以及其等可 ' 以直接地連接，或是經由肽連接子所連接的。當使用於本 文中，術語"連結"、”融合的”或是”融合”係可交換地使用。 此等術語係提及二個更多元素或組份藉由任何的手段（包 Q 括化學共軛或是重組手段)結合在一起。一"符合框架的融 合”係提及結合2或更多個開放閱讀框架以形成連續的更長 的開放閱讀框架，以維持原始的開放閱讀框架之正確的閱 讀框架的方式。因而，形成的重組型融合蛋白質為含有2或 更多個區段之一單一蛋白質，其等係對應於由原始的開放 閱讀框架所編碼的多肽（該等區段本質上通常不會如此結 合）。縱然被融合的區段處處之閱讀框架係因而被做成連續 的，該等區段可以實際上或是空間上由，舉例而言，符合 框架的連接子的序列予以分隔開的。一”連接子”的序列為 77 201038734 一系列的一或更多個的胺基酸，其等係將融合蛋白質内的 二個多狀編碼區域分隔開。 本發明係針對一組成物，其包含本發明的一或更多個 多肽以及生物上可接受的載體。 於一些具體例中，該組成物包括一生物上可接受的”賦 形劑”，其中該賦形劑為一組份，或是組份的混合物，其係 使用於本發明的一組成物中以提供該組成物所欲的特徵， 以及也包括載劑。''生物上可接受的”意指一化合物、材料、 組成物、鹽類，及/或劑量形式，其係落於合理的醫學判斷 的範疇内，適合於接觸活細胞的組織而於所欲的接觸期間 無過度的毒性、刺激、免疫反應，或是其他的疑難的併發 症、相稱於合理的利益/風險比。能使用各種各樣的賦形 劑。於一些具體例中，賦形劑可以是，但不限於：鹼性劑、 安定劑、抗氧化劑、黏著劑、分離劑、塗覆劑、外部相 組份、經控制的釋放組份、溶劑、界面活性劑、濕潤劑、 緩衝劑、填料、軟化劑，或其等之組合。除了本文中討論 的該等之外，賦形劑能包括賦形劑Remington: The Science and Practice of Pharmacy, 21st ed. (2005)中列出的，但不限 於該等。於本文中以特定的分類包含一賦形劑（例如，”溶 劑")係預期要闡釋而非限制賦形劑的角色。一特定的賦形 劑可以落於多重的分類之内。 本發明進一步有關本文中說明的多肽之任一者的一片 段、變異體、衍生物，或是類似物。 本發明的多肽可以是一重組型多肽、一天然的多肽， 78 201038734 或是一合成性多肽。 宿主細胞 本毛月係針對一宿主細胞，其表現以上說明的核酸分 子及重組㈣时子的任-者，以及料之組合。 ΟSSCO. 5% SDS at room temperature for 15 min, followed by 2X ssc ' 〇 5% SDS at 45 〇 c for 30 min, followed by 〇 2X ssc, 〇 5% 3] 〇 8 at 5 〇 0 (^ Repeat the second period of 3 〇minD for more severe conditions, at a higher temperature 0 line cleaning, the towel cleaning system is exactly the same as above, except for the last two 30 minutes to 〇·2χ ssc, 〇 The cleaning temperature of 5% Lizhong increased to C. The harsh conditions of the group are used at 65 ° C. IX SSC’ 0. 1/〇 SE) The second final cleaning in S. The extra set is highly rigorous.  The condition is defined as: at 65 ° Cf at 0. 1X SSC, 0. Hybridization in 1% SDS and 2X SSC, 〇. The l% SDS is then washed with χ χ ssc, 〇 1% sds. The present invention is directed to an isolated nucleic acid molecule comprising a multi-nuclear sequence which is fully complementary to any of the four multi-nuclear auditory phases of the above. The term "complementary" is the relationship between the acidity of this hybrid. For example, with respect to DNA, the glandular line is complementary to the thymus gland and the cytosine is complementary to guanosine. In some specific examples, the polynucleic acid or nucleic acid is DNA. In the case of DNA I, - nucleic acid A molecule comprising a polynucleotide encoding a polypeptide comprising a promoter and/or other transcriptional or translational control element operatively associated with one or more coding regions. The coding region of a gene product (eg, a polypeptide) is associated with - or more of the regulatory sequence of 59 201038734, in such a way that the expression of the gene product is under the influence of regulatory sequences or under the control of regulatory sequences Two DNA fragments (eg, a polypeptide coding region and a promoter associated with it) are "operably linked", if the induction of a promoter function results in the transcription of the mRNA encoding the desired gene product and The nature of the linkage between the two DNA fragments does not interfere with the ability of the expression-regulated sequence to indicate the expression of the gene product or the ability to interfere with the DNA template to be transcribed. Thus, the promoter region will be encoded with - The polynucleic acid sequence of the polypeptide is operatively associated, such that the promoter is capable of causing transcription of that polynucleotide sequence. The promoter may be a cell-specific promoter that is only indicative of the substantial transcription of the DNA within the predetermined cell. In general, a coding region is located at a promoter 3. The promoter may be derived from a natural gene, or a different element derived from a different promoter found in nature. , or even a synthetic DNA segment. As is known to those skilled in the art, different promoters can refer to genes that are not in different tissues or cell types, or at different stages of development, or are different. The expression of genes that respond to environmental or physiological conditions. Promoters, which cause a gene to behave most of the time, in most cell types, are often referred to as "constitutive promoters". Further recognition of regulation in most cases The exact boundaries of the sequences are not fully defined, and DNA fragments of different lengths can have the same promoter activity. A promoter is generally transcribed. The 3, the end of the position is bounded, and extends to the upstream (5, direction) to include the minimum number of bases or elements necessary for transcription of the detectable level above the starting background data. The _ transcriptional start position (suitably defined, for example, by nuclease S1) is found within 60 201038734 and the protein binding domain (consistent sequence) responsible for the binding of RNA polymerase. Suitable regulatory regions include The nucleic acid region is located upstream of a coding region (5' non-coding sequence), within a coding region, or downstream of a coding region (3' non-coding sequence), and transcription and RNA processing of the coding region that affects it. Whether it is stability or translation, the regulatory regions can include: promoters, translational leader sequences, RNA processing sites, effector binding sites, and stem-loop structures. In addition to a promoter, other transcriptional control elements, such as enhancers, operators, repressors, and transcription termination signals, can be operatively associated with the polynucleotide to indicate cell-specific transcription. The boundaries of this coding region are determined by the start codon at the 5' (amino) terminus and the translation stop codon at the 3' (wei) terminus. A coding region can include, but is not limited to, a prokaryotic region, a cDNA derived from mRNA, a genomic DNA molecule, a synthetic DNA molecule, or an RNA molecule. If the coding region is expected to be expressed in an eukaryotic cell, a polyadenylation signal and a transcription termination sequence will usually be located in the coding region. q In certain aspects of the invention, polynucleotide sequences having at least 20 bases, at least 30 bases, or at least 50 bases and hybridized to the polynucleotide sequences of the invention can be used as PCR Introduction. Typically, in PCR-amplified techniques, the primers have different sequences and are complementary to each other. The sequence of such primers should be designed to provide efficient and faithful replication of the target nucleic acid, depending on the desired test conditions. Methods for designing PCR primers are common and well known in the art. In general, the two short segments of the sequence can be used in a polymerase chain reaction (PCR) procedure to amplify longer nucleic acid fragments encoding homologous genes from DNA or RNA 61 201038734. The polymerase chain reaction can also be performed in a library of selected nucleic acid fragments, wherein the sequence of one primer is derived from the nucleic acid fragment and the sequence of the other primer is utilized to the coding microorganism by the presence of a polyadenylation bundle The 3' end of the mRNA precursor of the gene. Optionally, the sequence of the second primer can be based on the sequence derived from the selection vector. In addition, specific primers can be designed and used to amplify portions or full length of the sequence. The resulting amplification product can be directly labeled during the amplification reaction or labeled after the amplification reaction, and used as a probe to isolate the full length DNA fragment under suitable stringent conditions. Thus, the nucleic acid molecules of the invention can be used to isolate genes which encode homologous proteins from species of the same or other species or bacteria. The singularity of homologous genes using sequence dependent processes is well known in the art. Examples of sequence-dependent processes include, but are not limited to, methods of hybridization of nucleic acids, and methods of DNA and RNA amplification, as exemplified by various techniques for nucleic acid amplification (eg, polymerase linkage) Reaction, Mullis et al, U.S. Patent No. 4,683,202; ligase chain reaction (LCR) (Tabor, S. Etc., Proc.  Acad·Sci· USA 82: 1074 (1985)); or stock replacement amplification (SDA; Walker et al., Proc.  Natl.  Acad.  Sci.  U. S. A.  89: 392 (1992)) In some embodiments, the isolated nucleic acid molecules of the invention are used to isolate nucleic acid molecules from homologous organisms from other organisms to identify similar or improved PUFA forms. PUFA synthase. In some embodiments, the isolated nucleic acid molecules of the invention are used to isolate nucleic acid molecules from homologous organisms from other organisms, which are involved in the production of high amounts of DHA. 62 201038734 The nucleic acid molecule of the present invention also comprises a polynucleotide sequence encoding a domain of a PUFA synthase gene, a PUFA synthase gene, or a fragment of a PUFA synthase gene fused to a marker sequence in frame, the marker sequence The detection of the polypeptide of the invention is permitted. The sequence of markers includes auxotrophic or dominant markers known to those skilled in the art, such as: ZEO (Zeocin), NEO (G418), hygromycin, Shi Shenhua, HPH, NAT, and the like. The invention also encompasses variants of the PUFA synthase gene. Variants may contain coding regions, non-coding regions, or changes within the two. Examples of variants of a polynucleotide sequence contain alterations that result in silent substitution, addition, or deletion, but do not alter the nature or activity of the encoded polypeptide. In some embodiments, the polynucleotide sequence variation system is produced by silent substitution due to the degeneracy of the genetic code. In further embodiments, polynucleotide sequence variants can be produced for a variety of reasons, for example, to optimize the expression of a codon for a particular host (eg, to change the Thraustochytrium m RN A The codons within it become preferred by other organisms (e.g., E. coli or steer vesev/s/ae). Dual gene variants, orthologs, and/or homologs are also provided in the present invention. Genes, dual gene variants, splicing variants, full length coding portions, orthologs of the genes described herein can be obtained using procedures known in the art, using information disclosed herein. And / or a homologue. For example, dual gene variants and/or species homologs can be isolated and identified by making appropriate probes or primers from the sequences provided herein, as well as for screening for dual gene variants 63 201038734 And/or an appropriate source of nucleic acid for the desired homolog. The present invention is directed to a recombinant nucleic acid molecule comprising: any one of the nucleic acid molecules described above or a combination thereof, and a transcriptional control sequence. In some embodiments, the recombinant nucleic acid molecule is a recombinant plastic carrier. The present invention is directed to a method of making a recombinant vector' which comprises inserting one or more singly isolated nucleic acid molecules as described herein into a vector. The vector of the present invention, for example, can be a selection vector or a expression vector. The vector may be, for example, in the form of a plastid, a viral particle, a bacteriophage or the like. The polynucleotide sequences of the present invention can be included in any of a wide variety of expression vectors for expressing a polypeptide. Such vectors include chromosomes, non-chromosomes, and synthetic DNA or RNA sequences, for example, derivatives of SV40; bacterial plastids; and yeast plastids. However, any other suitable carrier known to those skilled in the art can be used. Suitable DNA sequences can be inserted into the vector by a variety of procedures. In general, the DNA sequence is inserted into a suitable restriction endogenous nuclease, as known in the art, and the remainder is considered to be within the scope of those skilled in the art. Such Procedures The present invention also encompasses recombinant constructs comprising a plurality of polynucleotide sequences as described above, the constructs comprising a set of bodies, for example or alternatively, a _ insert 2::: In the sample 64 201038734, the construct further comprises a sequence operatively associated with the regulation of the sequence 'including, for example, a promoter. A large number of suitable vectors and promoters are also known to the artist and are commercially available. Polypeptides The present invention is directed to isolated polypeptides comprising a PUFA synthase protein and a region of amino acid sequences derived from isolated microorganisms deposited under ATCC accession numbers ρτΑ_9695 and ΡΤΑ-10212. 0 As used herein, the term "polypeptide" is intended to include a single, conjugated peptide and plural polypeptides, and is referred to as being linked linearly via a guanamine bond (also known as a peptide bond). a molecule consisting of a monomer (amino acid). The S-Peptide's reference to any chain of 2 or more amino acids, and not to a specific length of product. Thus, a peptide, a dipeptide, a quinone peptide, an oligopeptide, a "protein", an "amino acid chain" or a chain or a chain (also referred to as (10)) of 2 or more amino acids is used. Any other term is included in the definition of "polypeptide", and the term "polypeptide" can be used in place of or in exchange for any of these terms. The term "polypeptide" is also intended to refer to the product of post-expression modification of the polypeptide, privately, without limitation: saccharification, acetylation, phosphorylation, guanidation, by known protecting groups/blocking groups Derivatization of 'proteolytic breakdown, or modification by the naturally occurring amino acid of the well. A polypeptide as described herein may include fragments, variants thereof, and derivatives thereof, without limitation. The terms "fragment variant", "derivative" and analog" when referring to a polypeptide include any polypeptide that retains the activity of at least some organisms. The polypeptide fragment can include: a proteolytic fragment, a fragment of the J clip 65 201038734, and a fragment that more readily reaches the site of action when delivered to an animal. The polypeptide fragment further includes any portion of the polypeptide comprising the antigenic or immunological epitope of the native polypeptide, including lines, and three-dimensional epitopes. The polypeptide fragment can comprise a variant region comprising a fragment as described above, and also a polypeptide having an amino acid sequence which is altered by amino acid substitution, deletion, or insertion. Variants can occur naturally, such as a pair of even gene variants. The "dual gene variant" is an optional form of the expected gene that occupies a given position on the chromosome of an organism. Non-naturally occurring variants can be produced using mutagenic techniques known in the art. The polypeptide fragments of the invention can comprise a conservative or non-conservative amino acid substitution, scrutiny, or addition. Variant polypeptides may also be referred to herein as "polypeptide analogs." The polypeptide fragments of the invention can also include a derivative molecule. As used herein, a ''derivative' of a polypeptide or a polypeptide fragment refers to a subject polypeptide having one or more residues chemically derivatized by a reaction of a functional side group. . "Derivatives" also include such peptides, which contain one or more of the 20 standard amino acids, or a naturally occurring amino acid derivative. For example, 4-hydroxyproline can be substituted for indoleamine. Acid; 5-hydroxyisamino acid can be substituted for lysine; 3-mercapto histidine can replace histidine; serine can replace serine; and ornithine can be substituted for lysine. A polypeptide can be encoded by any of the nucleic acid molecules of the invention. The invention is directed to an isolated polypeptide comprising Pfalp (SEQ ID NO: 2 or Sequence ID: 69), Pfa2p (SEQ ID NO: :4 or sequence identification number: 71), Pfa3p (sequence identification number: 6 or sequence identification 66 201038734 number: 73), and the amino acid sequence of the combination thereof has an identity of at least 8 〇. An acid sequence, wherein the polypeptides comprise one or more PUFA synthase activities. The invention is directed to a polypeptide comprising at least 8 amino acid sequences in the field of one or more PUFA synthase enzymes of the PUFA synthase of the invention. 〇% identity of the amino acid sequence. In the embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to an amino acid sequence in pfal 0 (SEQ ID NO: 2 or SEQ ID NO: 69), comprising one or more Multiple puFA synthesis_domains. In some specific examples, the polypeptide is comprised with Pfal ( sequence identification.  The amino acid sequence of No. 2 or Sequence Identification Number: 69) having at least 80% identity of amino acid sequence, comprising one or more puFA synthetase domains, for example: ks domain (SEQ ID NO: 8 Or sequence identification number: 75), MAT field (sequence identification number: 1〇 or sequence identification number: 77), ACp field (eg. Sequence identification number: any one of 14, 16, 18, 20, 22, 24, 81, Q 83, 85, 87, 89, 91, 93, 95, 97 or 99), 2 or more ACP fields Combination, for example: 2, 3, 4, 5, 6, 7, 8, 9 or 10 ACP fields, including tandem fields (sequence identification number: 12 or sequence identification number: 79, and parts thereof), KR Field (sequence identification number: 26 or sequence identification number: 101), DH field (sequence identification number: 28 or sequence identification number: 119), and combinations thereof. In some embodiments, the polypeptide comprises two or more amino acid sequences, wherein each of the at least two or more amino acid sequences is Pfalp (SEQ ID NO: 2 or Sequence ID: 69 The amino acid sequence in the ) is 80% identical and comprises one or 67 201038734 more PUFA synthetase domains. In some embodiments, the at least two or more amino acid sequences are 80% identical to the same amino acid sequence in Pfalp (SEQ ID NO: 2 or SEQ ID NO: 69). More areas of PUFA synthase. In some embodiments, the at least two or more amino acid sequences are 80% identical to the amino acid sequence different from Pfalp (SEQ ID NO: 2 or SEQ ID NO: 69), each of which comprises _ or more PUFA synthetase fields. In some embodiments, the at least two or more amino acid sequences are 80% identical to the amino acid sequence different from Pfalp (SEQ ID NO: 2 or Sequence ID: 69), wherein The level of the corresponding field in Pfalp (SEQ ID NO: 2 or Sequence Identification Number: 69) 'The at least 2 or more amino acid sequences are located at the same level or different levels in the polypeptide. In some embodiments, the at least two or more amino acid sequences are 80% identical to the amino acid sequence in Pfalp (SEQ ID NO: 2 or Sequence ID: 69), comprising one or more Multiple PUFA synthetase domains' eg Ks domain (sequence identification number: 8 or sequence identification number: 75), MAT domain (sequence identification number: 1〇 or sequence identification number: 77) 'ACP domain (eg sequence identification number) : any of the 14, 14, 18, 20'22, 24, 8 83, 85, 87, 89, 91, 93, 95, 97 or 99), 2, 3, 4, 5, 6, 78, Combination of 9 or 1 () heart fields, including tandem fields (sequence identification number: 12 or sequence identification number: 79, and parts thereof), KR field (sequence identification number ^ or sequence identification number) G1), DH Field (sequence identification number: a sequence identification number and a combination thereof, etc. In some specific examples, the polypeptide comprises 丄68 201038734 or more in PfaiP (sequence identification number: 2 or sequence identification number: 69) Amino acid sequence 'includes - or more PUFA synthetase fields, including any individual field Combination of one or more copies with one or more copies of any other individual field. In some embodiments, the invention is directed to a polypeptide comprising: with pfa2p (sequence recognition, ": 4 or contact: Chuan The silk acid sequence of the towel has a homologous amino acid sequence of >, 80%, and contains - or more puFA synthetase fields. In some specific examples, the nucleus comprises an amine group in Pfa2p. The amino acid sequence (sequence identification number: 4 or sequence identification number: 71) has at least identity amino acid sequence, including one or more puFA synthetase fields, for example: KS domain (sequence identification number: 3〇 or Sequence identification number: 103), CLF field (sequence identification number: 32 or sequence identification number: 1〇5), AT field (sequence identification number: 34 or sequence identification number: 1〇7), ER field (sequence identification number: 36 or a sequence identification number: 1〇9), and combinations thereof, etc. In some embodiments, the polypeptide comprises 2 or more amino acid sequences 'where the at least 2 or more amino acid sequences Each line with pfa2p (sequence identification number: 4 or sequence identification) The amino acid sequence in No. 71) is 80% identical and comprises one or more PUFA synthetase domains. In some embodiments, the at least two or more amino acid sequences are associated with Pfa2p (sequence Identification number: 4 or sequence identification number: 71) The same amino acid sequence has 80 〇 / 〇 identity. In some specific examples, the at least 2 or more amino acid sequence and Pfa2p (sequence identification The different amino acid sequences in the number: 4 or sequence identification number: 71) are 80% identical, each comprising one or more PUFA synthetase domains. In some embodiments, the at least two or more amino acid sequences differ from Pfa2p (SEQ ID NO: 4 or Sequence ID: 71) by a 69 201038734 amino acid sequence of 80 Å. The identity, which when comparing pfa2p (sequence identification editing. 4 or the sequence of the corresponding domain in the sequence identification number: 71), the J 2 or more amino acid sequences are located at the same level or different levels in the polypeptide. In some embodiments, the at least two or more amino acid sequences are 80% identical to the amino acid sequence in Pfa2p (SEQ ID NO: 4 or Sequence ID: 71), comprising one or more Multiple puFA synthetase fields, for example: KS domain (sequence identification number: 3〇 or sequence identification number: 1〇3), CLF domain (sequence identification number: 32 or sequence identification number h〇5), AT domain (sequence identification) No.: 34 or sequence identification number: 1〇7), er field (sequence identification number: 36 or sequence identification number: 1〇9), and combinations thereof. In some embodiments, the polypeptide comprises one or more amino acid sequences in Pfa2p (SEQ ID NO: 4 or Sequence ID: 71), comprising one or more PUFA synthetase domains, including any A combination of one or more copies of an individual field with one or more copies of any other individual field. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to an amino acid sequence in Pfa3p (SEQ ID NO: 6 or SEQ ID NO: 73), comprising one More or more puFA synthetase fields. In some embodiments, the polypeptide comprises an amino acid sequence having at least 8% homology to the amino acid sequence in pfa3p (SEQ ID NO: 6 or SEQ ID NO: 73), comprising - or more The field of PUFA synthetase, for example: DH domain (eg: sequence identification number: 38, sequence identification number: 40, sequence identification number: 111 ' or sequence identification number u13), er field (sequence identification number: 42 or sequence identification number) :115), and its group of 70 201038734. In some embodiments, the polypeptide comprises two or more amino acid sequences, wherein each of the at least two or more amino acid sequences is Pfa3p (SEQ ID NO: 6 or Sequence ID: 73 The amino acid sequence in the) is 80% identical and comprises one or more domains of PUFA synthase. In some embodiments, the at least two or more amino acid sequences are 80% identical to the same amino acid sequence in pfa3p (SEQ ID NO: 6 or SEQ ID NO: 73), comprising one or More areas of PUFA synthase. In some embodiments, the at least two or more amino acid sequences are 80% identical to the amino acid sequence different from Pfa3p (SEQ ID NO: 6 or SEQ ID NO: 73), each of which comprises One or more fields of PUFA synthetase. In some embodiments, the at least two or more amino acid sequences are 8% identical to the amino acid sequence different from pfa3p (SEQ ID NO: ό or SEQ ID NO: 73). When comparing the ranks of the corresponding domains in pfa3P (SEQ ID NO: 6 or Sequence ID: 73), the at least two or more amino acid sequences are at the same level or different grades in the polypeptide. In some embodiments, the at least two or more amino acid sequences are 80% identical to the amino acid sequence in Pfa3p (SEQ ID NO: 6 or SEQ ID NO: 73), comprising - or Multiple pUFA synthetase domains, for example: DH fields (eg. Sequence identification number: 38 'sequence identification number: 40, sequence identification number. Ill, or sequence identification number: 113), ER field (sequence identification number: 42 or sequence identification number: 115), and combinations thereof. In some embodiments, the side polypeptide comprises one or more amino acid sequences in Pfa3p (SEQ ID NO: 6 or Sequence ID: 73), comprising one or more PUFA synthesis_7 member domains, Includes one or more copies of any individual field and any combination of any of the other individual fields or multiple copies of 71 201038734. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to Sequence Identification Number 2: or Sequence Identification Number: 69, wherein the polypeptide comprises a moiety selected from the group consisting of The PUFA synthase activity of the resulting group: KS activity, MAT activity, ACP activity, KR activity, DH activity, and combinations thereof. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 8 or sequence identification number: 75, wherein the polypeptide comprises KS activity. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 10 or sequence identification number: 77, wherein the polypeptide comprises MAT activity. In some embodiments, the invention is directed to a polypeptide comprising and sequence identification numbers: 14, 16, 18, 20, 22, 24, 81, 83, 85, 87, 89, 91, 93, 95, 97 Or any one of 99 having at least 80% identity amino acid sequence, wherein the polypeptide comprises ACP activity. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 12 or sequence identification number: 79, wherein the polypeptide comprises ACP activity. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to an amino acid sequence within sequence identification number: 12, wherein the polypeptide comprises ACP activity. In some embodiments, the amino acid sequence is at least 80% identical to the amino acid sequence of Sequence Identification Number: 12, which encodes 1, 2, 3, 4, 5 or 6 ACP domains, 72 201038734 The polypeptide comprises an ap activity associated with one or more ACP domains. Sequence identification numbers: 14, 16, 18, 20, 22, and 24 are representative amino acid sequences comprising: a single ACP domain in sequence identification number: 12. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to an amino acid sequence within sequence identification number: 79, wherein the polypeptide comprises ACP activity. In some embodiments, the amino acid sequence has at least 80% identity to the amino acid sequence of Sequence Identification Number: 79, which comprises 1, 2, 3, 4, 5, 6, and 7, 8, 9 or 1 ACP domain, wherein the polypeptide comprises ACP activity associated with one or more ACP domains. Sequence Identification Numbers: 81, 83, 85, 87, 89, 91, 93, 95, 97, and 99 are representative amino acid sequences comprising: Sequence Identification 'Number: 79 in a single ACP domain. In some embodiments, the invention is directed to a polypeptide comprising a sequence identification number. 26 or sequence identification number: 1 amino acid sequence having at least 8% identity, wherein the polypeptide comprises KR activity. ☆ ☆ In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 8% homology to the sequence identification number: 28 or SEQ ID NO: 119, wherein the polypeptide comprises Dh activity. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence of the same nature as the sequence identification number: 4 or the sequence number: 71, wherein the polypeptide comprises a moiety selected from the group consisting of The group of PUFA synthase AI-mailing, CLF^, at activity, er activity, and combinations thereof. In some embodiments, the invention is slanted in a polymorphic form comprising an amino acid sequence having at least 80% identity to sequence 73 201038734, column identification number: 30 or sequence identification number: 103, wherein the polypeptide comprises KS active. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 32 or sequence identification number: 105, wherein the polypeptide comprises CLF activity. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 34 or sequence identification number: 107, wherein the polypeptide comprises AT activity. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 36 or sequence identification number: 109, wherein the polypeptide comprises ER activity. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 6 or sequence identification number: 73, wherein the polypeptide comprises a moiety selected from the group consisting of The PUFA synthase activity of the resulting group: DH activity, ER activity, and combinations thereof. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 38, wherein the polypeptide comprises DH activity. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to the sequence identification number: 40, wherein the polypeptide comprises DH activity. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to the sequence identification number: 111, wherein the polypeptide comprises DH activity. 74 201038734 In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to the sequence identification number: 113, wherein the polypeptide comprises DH activity. In some embodiments, the invention is directed to a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 42 or sequence identification number: 115, wherein the polypeptide comprises ER activity. In some embodiments, the polypeptides comprise an amino acid sequence at least about 80%, 85%, or 90% identical to the amino acid sequence reported herein, or comprise the amine group as reported herein. The acid sequence has at least about 95%, 96%, 97%, 98%, 99%, or 100% identity of the amino acid sequence. With respect to a polypeptide', it has, for example, at least 95°/in relation to an interrogated amino acid sequence of the invention. "identical," amino acid sequence" is meant to mean that the amino acid sequence of the subject polypeptide is identical to the amino acid sequence of the query, except for the amino acid sequence in question. The subject polypeptide sequence in each of the 1 amino acid groups can include up to 5 amino acid differences. In other words, 'to obtain a polymorphism, which has at least 95% homology to the amino acid sequence of the query, and up to 5% of the amino acid residues in the subject polypeptide can be inserted. Loss (indds) or replaced with another amino acid. Such alterations in the reference sequence can occur at the amino or carboxy terminus of the reference amino acid sequence or at any place between the terminal positions 'individually dispersed in the residue of the reference sequence towel Within one or more of the linked groups of the reference sequence. In practice, whether any particular polypeptide has an amine that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to the amine 75 201038734 acid sequence of the present invention. The base acid sequence, for example, can be routinely determined using known computer programs as discussed above. As discussed above, a method for determining the best overall match between an interrogation sequence (a sequence of the invention) and a subject sequence can be determined using the alignment of the sequences and the calculation of the identity score. The arrangement is performed using the computer program AlignX, which comes from Invitrogen (www. Invitrogen. Com) carrier NTI Suite 10. 0 sets of software components. The arrangement is arranged using ClustalW (J.  Thompson et al., Nucleic Acids Res.  22(22): 4673-4680 (1994) to perform. Use the system pre-designed sub-matrix Blosum62mt2. The system presets the space opening penalty to be 10 and the space extension penalty is 0. 1. In a further aspect of the invention, the nucleic acid molecule, etc., is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequences disclosed herein. A polynucleotide sequence encoding a polypeptide having one or more PUFA synthase activities. A polypeptide having one or more PUFA synthase actives exhibits one or more activities which are similar to the activity of one or more of the PUFA synthase of the present invention, but are not necessarily related to the PUFA synthase of the present invention One or more of the activities are identical. Of course, due to the degeneracy of the genetic code, those skilled in the art will immediately recognize that most of the nucleic acid molecules have at least 80 Å/°, 85°/ of the polynucleotide sequence described herein. 90. A polynucleotide sequence of /〇, 95%, 96%, 97%, 98%, or 99% identity, encoding a polypeptide having a PUFA synthase functional activity". In fact 'so all of the degenerate variants of any of the polynucleotide sequences encode the same polypeptide, in many cases, it can be replaced by conservation, and conservational functionality by familiarity with this artist 76 201038734 In the field, the polypeptide will exhibit activity and the knowledge will be predicted. In certain aspects of the invention, the polypeptides and polynucleotides of the invention are provided in isolated form, e.g., purified to homogeneity. Alternatively, the polypeptides and polynucleotides of the present invention can be produced synthetically by conventional synthesizers. "Similarity" between two polypeptides as known in the art is provided by comparing the amino acid sequence of the amino acid sequence and the sequence of a second polypeptide to a conserved amino acid substituent of the polypeptide Decide. In some embodiments, the polypeptide of the invention is a fusion polypeptide. As used herein, a π-fusion polypeptide '' means a polypeptide comprising a first polypeptide that is linearly linked to a second polypeptide via a peptide bond. The first 'polypeptide and the second polypeptide may be the same or different, and the like may be directly linked or linked via a peptide linker. As used herein, the terms "linked", "fused" or "fused" are used interchangeably. These terms refer to two more elements or components that are combined by any means (including chemical conjugation or recombination). A "conformance to framework" refers to the way in which two or more open reading frames are combined to form a continuous longer open reading frame to maintain the correct reading frame of the original open reading frame. A recombinant fusion protein is a single protein containing one or more of the segments, which corresponds to the polypeptide encoded by the original open reading frame (the segments are not normally so integrated in nature). The reading frame at each section is thus made continuous, and the segments can be separated, in fact, or spatially by, for example, a sequence of linkers conforming to the frame. A "linker" The sequence is 77 201038734 a series of one or more amino acids separated by two polymorphic coding regions within the fusion protein. The present invention is directed to a composition comprising one of the present invention Or more polypeptides and a biologically acceptable carrier. In some embodiments, the composition comprises a biologically acceptable "excipient" wherein the excipient is a component Or a mixture of components, which is used in a composition of the invention to provide the desired characteristics of the composition, and also includes a carrier. ''Biologically acceptable'" means a compound, material, composition The substance, salt, and/or dosage form is within the scope of sound medical judgment and is suitable for contact with living tissue without excessive toxicity, irritation, immune response, or other during the desired exposure. The complication of the problem is commensurate with a reasonable benefit/risk ratio. A wide variety of excipients can be used. In some embodiments, the excipient can be, but is not limited to, an alkaline agent, a stabilizer, an antioxidant, an adhesive, a separating agent, a coating agent, an external phase component, a controlled release component, a solvent, a surfactant, a wetting agent, a buffer, a filler, a softener, or a combination thereof. In addition to those discussed herein, excipients can include excipients Remington: The Science and Practice of Pharmacy, 21st ed.  Listed in (2005), but not limited to such. The inclusion of an excipient (e.g., "solvent" in a particular class herein is intended to be illustrative and not limiting the role of the excipient. A particular excipient may fall within multiple sub-categories. Further a fragment, variant, derivative, or analog of any of the polypeptides described herein. The polypeptide of the invention may be a recombinant polypeptide, a natural polypeptide, 78 201038734 or a synthetic polypeptide. The host cell is a host cell that exhibits the nucleic acid molecules described above and any of the recombinant (iv) time, and combinations of materials.

田使用於本文巾’術語"表現"係提及—基因生產生物 化子物(舉例* § ’⑽八或是多肽)的過程。該過程包括該基 因於細胞内之功能性存在的任何的展示，包括，沒有限制， 基因剔除’以及暫時性表現以及和且安定的表現二者。其 包括，’又有限制，該基因的轉錄成信使RNA(mRNA)、轉移 RNA(tRNA)、小髮夾式RNA (雜獻）、小干擾性 RNA(siRNA) ’或是任何其他的RNA產物，以及此（等)mRNA 轉譯成多狀。設若最終所欲的產物為生物化學物，表現會 包括生物化學物以及任何的前驅物之創造。 為了產生一或更多個所欲的多不飽和脂肪酸，一種宿 主細胞能經基因修飾以將本發明的PUFA合成酶系統引入 至該宿主細胞中。 當基因修飾有機體以表現如本發明的一 PUFA合成酶 系統時，一些宿主有機體可内源地表現需要的輔助蛋白質 以及一PUFA合成酶系統俾以產生PUFAs。然而，用編碼一 或更多個輔助蛋白質之核酸分子來轉形一些有機體俾以使 得該有機體生產PUFAs或是提升該有機體之PUFAs的生產 可能是必須的，即使該有機體内源性地產生一同源性的輔 助蛋白質。一些異源性輔助蛋白質能比該宿主細胞的内源 性輔助蛋白質更有效地或是有效率地運作該等經轉形的 79 201038734 PUFA合成酶蛋白。 輔助蛋白質於本文中定義為不被視為該核心PUFA合 成酶系統的部分（亦即，非PUFA合成酶酵素複合體本身的 部分）之蛋白質，但是其等對於使用本發明的核心PUFA合 成酶酵素複合體之PUFA的生產或有效PUFA的生產可以是 必須的。舉例而言，為了產生PUFAs，一PUFA合成酶系統 必須與會自辅酶A轉移4’-磷酸泛硫醇部分至醯基載體蛋白 質(ACP)領域之輔助蛋白質工作。因而，一puFA合成酶系 統能被視為包括至少一個4'-磷酸泛硫醇轉移酶(ppTase)領 域，或此一領域能被視為該PUFA合成酶系統之輔助領域或 蛋白質。PPTases之結構與功能性特徵已經詳細描述，例 如，於美國申請公開案第2〇〇2/0194641號；第2004/0235127 號；以及第20〇5/〇1〇〇995號之中。 具有4 -磷酸泛硫醇轉移酶(ppTase)生物活性（功能）之 領域或蛋白質係特徵為該酵素會自輔酶A轉移4，_碟酸泛硫 醇。P刀至4醯基載體蛋白質(Acp)。此轉移至Acp之無變化 的絲胺酸會活化未活化之不完全形式至完全形式 。在聚酮 ,δλ a t SlL^(phosphopantetheine group) 會與成長巾之醯錢形成麵。ρρτ_為—酵素家族，其 已於月曰肪酸口成、聚酮合成，以及非核酶體胜肽合成步驟 中充分地定特徵。許多PPTases序列為已知的，並已決定其 結晶結構(例如’ R峨r K.等人，EMBO L 18(23)細_31 (1999)) α及已鐘定出對於活性相當重要之胺基酸殘基突 變分析（Mofid M.R·箄人 D· u · 寸入，Biochemistry 43(14):4128-30 80 201038734 (2004))。 一異源性PPTase，其先前已經顯示出可辨識出裂殖壺 菌ACP領域作為受質，為念珠藻屬物種(TVoWoc印.)PCC 7120(先稱之為念珠藻屬物種w.) pec 712〇) 之Hetl蛋白質。Hetl係存在於念珠藻屬基因組中，已知負 責長鏈羥基-脂肪酸的合成，其為該有機體的異形細胞存在 之醣脂層的一成分（Black and Wolk，J. Bacteriol. 176: 2282-2292 (1994) ; Campbell等人，Arch. Microbiol. 167: 251-258 (1997))〇HetI似乎可活化該基因組中存在之一蛋白 質，HglE之ACP區塊。含有Hetl之序列與建構物已經說明 於，例如，美國申請公開案第2007/0244192之中號，以其 之整體併入本文中作為參考資料。 先前已經展現出可辨識裂殖壺菌ACP領域之另一異源 性PPTase為Sfp，其係自枯草桿菌（Bacillus subtilis)所衍生。 Sfp已經被充分地定特徵並廣泛使用，由於其可辨識廣範圍 受質之能力。根據已公開之序列資訊（Nakana,等人， Molecular and General Genetics 232: 313-321 (1992))，先前 用於Sfp表現之載體，其係藉由選殖該編碼區域，與經定義 之上游-與下游側邊DNA序列，至pACYC-184選殖載體上而 製備。此建構物編碼一功能性PPTase，其展現出在適當條 件下可與裂殖壺菌Orfs於大腸桿菌内共表現，導致DHA累 積於這些細胞中的能力（參見，美國申請公開案第 2004/0235127號，以其之整體併入本文中作為參考資料）。 宿主細胞能包括：微生物細胞、動物細胞、植物細胞， 81 201038734 以及昆蟲細胞。合適的宿主的代表性實例包括細菌細胞； 嗜熱性或嗜溫性(mesophUc)細菌；海洋細菌；破囊壺菌； 真菌細胞，例如酵母菌；植物細胞；昆蟲細胞；以及經單 離的動物細胞。宿主細胞可以是未經轉染的細胞或是已經 用至少一種其他的重組型核酸分子轉染的細胞。宿主細胞 也能包括已經經改造以表現PUFA合成酶之基因轉殖細 胞。合適的宿主的挑選被視為落在熟悉此藝者從本文中的 教示之範轉内。 宿主細胞包括破囊壺菌目的任何微生物，例如：來自 一屬的微生物，包括’但不限於：破囊壺菌、拉比琳休洛 依迪斯菌（Ζί%咖认Μ/θ/ί/以）、加彭歐凱崔恩菌 ，以及此等屬範圍内的裂殖壺菌種包括， 但不限於：任何裂殖壺菌種’包括聚裂殖壺菌 limacimim)、微小裂殖壺菌（Schizochytrium mimitum) _，任何 破囊壺菌物種（包括先前的巫肯尼亞菌（t//A：em'a)物種，例 如：維蘇爾傑西斯巫肯尼亞菌（t/· 、阿墨巴戴亞 巫肯尼亞菌（i/· 、沙勒乃利阿納巫肯尼亞菌（^/ sarAraWima)、普羅弗達巫肯尼亞菌（C/· 、拉迪亞它 巫肯尼亞菌（t/_ rat//如a)'迷努它巫肯尼亞菌（f/ •⑽加)及巫 肯尼亞菌物種BP-5601) ’以及包括：紋狀破囊壺菌 (r/^awMoc/z声Wwm Wr/aiww)、金黄色破囊壺菌、羅斯破囊壺 菌（77^<3«1^0<^>^〜所，〇15似所）；以及任何加彭歐凱崔恩菌物 種。破囊3?函目的囷株包括’但不限於：裂殖壺菌種（S31) 82 201038734 (ATCC 20888);裂殖壺菌種(S8) (ATCC 20889);裂殖壺菌 種(LC-RM) (ATCC 18915);裂殖壺菌種(SR21);聚裂殖壺 菌（Goldstein et Belsky) (ATCC 28209);裂殖壺菌 (Schizochytrium limacinum)(Honda et Yokochi) (IFO 32693);破囊壺菌種(23B) (ATCC 20891);紋狀破囊壺菌 (Schneider) (ATCC 24473);金黄色破囊壺菌（Goldstein) (ATCC 34304);羅斯破囊壺菌（Goldstein) (ATCC 28210); 以及加彭歐凱崔恩菌物種(LI) (ATCC 28207)。基因修飾之 合適的宿主微生物之其他的實例包括，但不限於：酵母菌， 包括.啤酒酵母、嘉士伯酵母菌（Sacc/zarowyce·? 或是其他酵母菌，例如：念珠菌屬、脆壁克 魯維酵母少vero—ce·?)或其他真菌，舉例而言，絲狀真 菌’如麴菌屬〇^/^似7/似）、紅黴菌屬、青黴 菌等。細菌細胞亦可使用作為宿主。這包括大 腸桿菌，其可用於發酵製程中。任擇地，例如乳酸菌 物種或桿菌（5ac"/⑽)物種的宿主能使用作 為宿主。 植物宿主細胞包括，但不限於：任何高等植物，包括 雙子葉與早子葉植物一者’以及消耗性植物（consumable plant)，包括作物植物與使用其油類之植物。此等植物能包 括，舉例而言：油菜(canola)、大豆、油菜子(rapeseed)、亞 麻仁（linseed)、玉米、紅花（saff|〇wer)、葵花與煙草 (tobacco)。其他植物包括已知可製造用於醫藥試劑、調味 劑、營養劑、功此性食品成分或化妝品活性成分之化合 83 201038734The field used in this article 'terms' "performance" refers to the process of gene production of biomass (for example, § '(10) eight or polypeptide). This process includes any display of the gene's functional presence within the cell, including, without limitation, gene culling' as well as transient performance and both stable and stable performance. It includes, 'restricted, transcription of this gene into messenger RNA (mRNA), transfer RNA (tRNA), small hairpin RNA (missing), small interfering RNA (siRNA)' or any other RNA product And this (etc.) mRNA is translated into multiple shapes. If the final desired product is a biochemical, the performance will include the creation of biochemicals and any precursors. To produce one or more desired polyunsaturated fatty acids, a host cell can be genetically modified to introduce a PUFA synthase system of the invention into the host cell. When genetically modifying organisms to exhibit a PUFA synthase system as in the present invention, some host organisms can endogenously express the desired accessory protein as well as a PUFA synthase system to produce PUFAs. However, it may be necessary to use a nucleic acid molecule encoding one or more accessory proteins to transform some organisms such that the organism produces PUFAs or enhance the production of PUFAs of the organism, even if the organism is produced together in vivo. Source-assisted protein. Some heterologous helper proteins are capable of operating the transformed 79 201038734 PUFA synthase protein more efficiently or efficiently than the host cell's endogenous helper protein. A helper protein is defined herein as a protein that is not considered part of the core PUFA synthase system (ie, a portion of the non-PUFA synthetase enzyme complex itself), but which is for use with the core PUFA synthase enzyme of the present invention. The production of PUFAs of the composite or the production of effective PUFAs may be necessary. For example, in order to produce PUFAs, a PUFA synthase system must work with an accessory protein that would transfer a 4'-phosphothiol moiety from coenzyme A to a thiol-containing carrier protein (ACP) domain. Thus, a puFA synthase system can be considered to include at least one domain of 4'-phosphothioate transferase (ppTase), or this field can be considered as an auxiliary domain or protein of the PUFA synthase system. The structural and functional characteristics of PPTases have been described in detail, for example, in U.S. Patent Application Publication No. 2,2,091, 946, 421, No. 2004/0235, 127, and No. 20, 5/1, 995. A domain or protein line having a biological activity (function) of 4-phosphome panthenol transferase (ppTase) is characterized in that the enzyme is transferred from coenzyme A 4, oxalate panthenol. P-knife to 4-mercapto carrier protein (Acp). This unchanged transition to Acp will activate the inactivated incomplete form to the complete form. In the polyketone, δλ a t SlL^(phosphopantetheine group) will form a surface with the money of the growth towel. The ρρτ_ is a family of enzymes that have been sufficiently characterized in the synthesis of ruthenium, polyketone synthesis, and non-ribosomal peptide synthesis steps. Many PPTases sequences are known and have determined their crystal structure (eg 'R峨r K. et al., EMBO L 18(23) fine _31 (1999)) α and amines which have been determined to be important for activity Mutation analysis of basal acid residues (Mofid MR 箄人D·u · 入入, Biochemistry 43(14): 4128-30 80 201038734 (2004)). A heterologous PPTase, which has previously been shown to recognize the Schizochytrium ACP domain as a substrate for the Candida species (TVoWoc.) PCC 7120 (first known as the Noctually species w.) pec 712 〇) Hetl protein. Hetl is found in the genome of Nostoc, and is known to be responsible for the synthesis of long-chain hydroxy-fatty acids, which is a component of the glycolipid layer in the heterogeneous cells of the organism (Black and Wolk, J. Bacteriol. 176: 2282-2292 (1994); Campbell et al., Arch. Microbiol. 167: 251-258 (1997)) HetI seems to activate one of the proteins present in the genome, the ACP block of HglE. Sequences and constructs containing Hetl are described, for example, in U.S. Patent Application Publication No. 2007/0244, the entire disclosure of which is incorporated herein by reference. Another heterologous PPTase that has previously been shown to be identifiable in the ACP field of Schizochytrium is Sfp, which is derived from Bacillus subtilis. Sfp has been well characterized and widely used due to its ability to identify a wide range of substrates. According to the published sequence information (Nakana, et al, Molecular and General Genetics 232: 313-321 (1992)), a vector previously used for Sfp expression, by cultivating the coding region, and defined upstream - Prepared with the downstream flanking DNA sequence on the pACYC-184 selection vector. This construct encodes a functional PPTase that exhibits the ability to co-exist with Schizochytrium sinensis Orfs in E. coli under appropriate conditions, resulting in the ability of DHA to accumulate in these cells (see, US Application Publication No. 2004/0235127). No., which is incorporated herein by reference in its entirety. Host cells can include: microbial cells, animal cells, plant cells, 81 201038734 and insect cells. Representative examples of suitable hosts include bacterial cells; thermophilic or mesophilic (mesophUc) bacteria; marine bacteria; Thraustochytrium; fungal cells such as yeast; plant cells; insect cells; and isolated animal cells . The host cell can be an untransfected cell or a cell that has been transfected with at least one other recombinant nucleic acid molecule. Host cells can also include gene transgenic cells that have been engineered to express PUFA synthase. The selection of a suitable host is considered to fall within the teachings of those skilled in the art. The host cell includes any microorganism of the order Thraustochytrium, for example: a microorganism from a genus, including but not limited to: Thraustochytrium, Labrin hollywood (Ζί%咖Μ/θ/ί/ ,), and the schistosomiasis in the genus of these species include, but are not limited to, any schizophrenic species 'including the schizophrenia limacimim, and the micro-Schizochytrium (Schizochytrium mimitum) _, any Thraustochytrium species (including previous W. Kenyan (t//A:em'a) species, eg: Vesul Jess, Kenyan bacterium (t/·, Amo Badeia Kenyan bacterium (i/·, Shah Nellie, Kenyan bacterium (^/ sarAraWima), Profida kenya (C/·, Radian, Kenyan bacterium (t/_ rat// as a) 'Indonesia witch Kenyan bacteria (f / • (10) plus) and witch Kenyan species BP-5601) 'and including: Thraustochytrium (r/^awMoc / z sound Wwm Wr / aiww), golden yellow broken Chlamydia, Thraustochytrium sclerotium (77^<3«1^0<^>^~, 〇15like); and any Gabon Oxygen species. Alfalfa strains include, but are not limited to, Schizochytrium species (S31) 82 201038734 (ATCC 20888); Schizochytrium species (S8) (ATCC 20889); Schizochytrium species (LC-RM) (ATCC 18915) Schizochytrium (SR21); Goldstein et Belsky (ATCC 28209); Schizochytrium limacinum (Honda et Yokochi) (IFO 32693); Thraustochytrium (23B) (ATCC 20891); Schneider (ATCC 24473); Golden Stem (Goldstein) (ATCC 34304); Goldstein (ATCC 28210); and Gabon Eucalyptus species (LI) (ATCC 28207). Other examples of suitable host microorganisms for genetic modification include, but are not limited to, yeast, including. Saccharomyces cerevisiae, S. cerevisiae (Sacc/zarowyce®? Other yeasts, for example: Candida, Kluyveromyces cerevisiae vero-ce·?) or other fungi, for example, filamentous fungi such as genus 麴 / ^ / ^ like 7 / like), red Fungi, penicillium, etc. Bacterial cells can also be used as a host. This includes E. coli, which can be used in fermentation processes. Optionally, for example, lactic acid bacteria species The host of the Bacillus (5ac"/(10)) species can be used as a host. Plant host cells include, but are not limited to, any higher plant, including dicotyledonous and early cotyledon plants' and consumable plants, including crop plants and plants using their oils. Such plants can include, for example, canola, soybean, rapeseed, linseed, corn, saff|〇wer, sunflower and tobacco. Other plants include compounds that are known to be manufactured for use in pharmaceutical agents, flavoring agents, nutrients, functional food ingredients or cosmetic active ingredients. 83 201038734

物的該等植物，或該植物經基因工程改造以製造這些化 合物/試劑者的植物。因而，可選擇任何植物物種或植物 細胞。植物與植物細胞，以及由此生長或衍生的植物之 實例包括’但不限於：由油菜可獲得的植物與植物細胞 (蕪菁）；油菜栽培品種NQC02CNX12 (ATCC PTA-6011)、 NQC02CNX21 (ATCC PTA-6644)，和NQC02CNX25 (ATCC PTA-6012)，以及栽培品種、育種栽培品種，以及自油菜栽 培品種NQC02CNX12、NQC02CNX21，和NQC02CNX25所 衍生的植物部分(各別地參見，美國專利案第7,355,100號、 第7,456,340號’和第7,348,473號）；大豆（黃豆）；油菜子（芥 屬物種），亞麻仁/亞麻（亞麻子(Z/A7MW似/加/心/所况所)）；玉蜀 泰（玉米）（玉米（Zea mfljAs));紅花（紅花（Cari/zamw·? //«cior/wi))，凑化（向曰蔡(好⑽仙似)）；於草（於草 {Nicotiana tabacum)) ’ 阿紅伯齐〈Arabidopsis thaliana)、巴 西栗子（巴西栗子CSei/w/eiiz’a ejcce/犯)）；篦麻子（篦麻子 COwwMmi));椰子（椰子（Coc;胡荽（胡 妥（Cor/a«(irww ίαί/vwm))，棉花（棉物種; 花生（落花生(Arachis hypogaea));荷荷巴（希蒙得木黃楊 c/nVzewA));芥菜（芥屬物種和白芥（57 祕α));油椰子（油椰子(五/⑽·? gw—m));橄欖（橄棍（〇/如 ewrpaea))，米（稻saiz.va));南瓜（南瓜 wax/wa))，大麥（大麥（//ori/eww vw/g^re));小麥（小麥 (TVaei/cww iiesi/vwm))，以及浮萍（浮萍物種 •s/?·))。來自此等及其他的植物之植物品種可以被生產、被 84 201038734 挑選的，或是為了所欲的特徵而最佳化，例如以下或與以 下有關聯的，但不限於：種子產量、耐倒伏性、羽化、疾 病抗性或耐受性、成熟、晚季植物完整性、植物高度、裂 果抗性、容易植物轉形、油含量，或是油形態。植物品種 可以經由植物育種來挑選，例如：純種的育種、週期性的 篩選育種、異種交配及回交育種，以及，例如：標誌辅助 性育種及耕種的方法。參見，如，美國專利案第7,348,473 號。 動物細胞包括任何經單離的動物細胞。 本發明係針對一宿主細胞，其表現本發明的一或更多 個核酸分子或重組型核酸分子，包括載體。 本發明係針對一種製造重組型宿主細胞的方法，其包 含將一重組型載體引入至一宿主細胞中。 宿主細胞可以是以本發明的載體予以基因工程改造的 (轉導的或轉形的或轉染的），該載體可以是，舉例而言，一 選殖載體或一表現載體。該載體可以是，舉例而言，以一 質體、一病毒顆粒、一噬菌體等等的形式。該載體含有如 本文中說明的一多核苷酸序列，以及一合適的啟動子或控 制序列，可以用來轉形一合適的宿主以允許由該多核苷酸 編碼的多肽之表現。宿主細胞的基因修飾也能包括較佳或 最佳的宿主密碼子使用之基因最佳化。 該經改造的宿主細胞能於改質成合適於活化啟動子、 篩選轉形體，或是擴增本發明的基因之慣用的營養培養基 之中予以培養。培養條件，例如：溫度、pH，以及類似物， 85 201038734 為先前使用於該宿主細胞篩選表現的該等培養條件，以及 對於熟悉此藝者為明顯的。 於一些具體例中，本發明係針對基因修飾一植物或植 物的部分以表現本文中說明的一PUFA合成酶系統，其包括 至少核心PUFA合成酶酵素複合體。如本文中定義的"植物 的部分(part of a plant)”或，，植物的部分(piant part)”包括任 何植物的部分，例如，但不限於：種子（成熟與未成熟）、油、 花粉、胚胎、花、果實、幼芽、葉子、根、樹幹、外植片 體等。於一些具體例中，該經基因修飾的植物或植物的部 分生產一或更多個PUFAs ’例如：EPA、DHA、DPA(n-3或 n-6)、ARA、GLA、SDA、其他的PUFA，以及其等之組合。 並不知道植物内源性含有一PUFA合成酶系統；因而，本發 明的PUFA合成酶系統能使用來改造具有獨特的脂肪酸生 產能力之植物。於進一步的具體例中’植物或植物的部分 被進一步基因修飾以表現至少一個PUFA合成酶輔助蛋白 質（例如，PPTase)。於一些具體例中’該植物為油類種子植 物’其中該油類種子，及/或油類種子中之油類，係包含PUFA 合成酶系統製造之PUFA。於一些具體例中，該等經基因修 飾的植物、植物的部分、油類種子，及/或油類種子中之油 類含有可偵測量之至少一個PUFA，其為該PUFA合成酶系 統之產物。於進一步的具體例中，此等植物、植物的部分、 油類種子’及/或油類種子中之油類實質上可不含中間產物 或副產物，其並非引入的PUFA合成酶系統的主要PUFA之 產物’且其並非由野生型植物内的内源性FAS系統所天然地 86 201038734 產生的。雖然野生型植物經由FAS系統生產—些短鏈或中鏈 PUFAs，例如18個碳之PUFAs，但新的或額外的puFAs會於 植物、植物的部分、油類種子，及/或油類種子中之油類中 生產係為經本文中說明的PUFA合成酶系統基因修飾之結 果。 植物之基因修飾可使用典型的菌株發育及/或分子遺 傳技術達成。參見，美國申請公開案第2〇〇7/〇244192號。 生產基因轉殖植物的方法係熟悉此藝者知道的，其中一種 編碼所欲的胺基酸序列之重組型核酸分子係被併入至該植 物的基因體之中。舉例而言，病毒載體能使用來生產基因 轉殖植物，例如：藉由用一病毒載體之單子葉的植物之轉 形’其係使用美國專利案第號5,569,597號；第5,589,367號； 和第5,316,931號之中說明的方法。藉由轉形來基因工程改 造或修飾植物的方法亦為本技藝所熟知的，包括生物性與 物理性轉形流程。參見，例如’ B.L. Miki等人，Procedures for Introducing Foreign DNA into Plants, in METHODS IN PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY 67-88 (Glick, B. R. and Thompson, J. E. eds., CRC Press, Inc.，Boca Raton, 1993)。此外，用於植物細胞或組織轉形與 植物再生之載體與體外培養方法為可獲得的。參見，例如， Μ· Y. Gruber等人，Vectors for Plant Transformation, in METHODS IN PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY 89-119 (Glick, B. R. and Thompson, J. E. eds., CRC Press, Inc_，Boca Raton, 1993)。 87 201038734 廣泛使用之用於引入一表現載體至植物中之方法係根 據農桿菌（Agrobacterium)之天然轉形系統。參見，例如， Horsch 等人，Science 227:1229 (1985)與美國專利案第 6,〇51，757號。根瘤農桿菌(A. tumefaciens)與髮根農桿菌（A. rhizogenes)為植物病理土壤細菌，其可使植物細胞基因轉 形。根瘤農桿菌與髮根農桿菌之Ti與Ri質體係各別攜帶使 植物基因轉形之對應基因。參見，例如，Kado, C. I., Crit. Rev.Plant. Sci· 10:1 (1991)。關於農桿菌載體系統與農桿菌 媒介之基因轉移方法之描述係提供於數種文獻中，包括 Gruber等人，如上所述；Miki等人，如上所述；Moloney等 人，Plant Cell Reports 8:238 (1989);美國專利案第 5，177,010 號；第 5，104,310號；第 5，149,645 號；第 5,469,976號；第 5,464J63號；第 4,940,838號；第 4,693,976號；第 5,591,616 號；第 5,231,019 號；第 5,463,174號；第 4,762,785 號；第 5,004,803號；和第5,159,135號；以及歐洲專利申請案第 0131624號、第 120516號、第 159418號、第 176112號、第 116718號、第 290799號、第 320500號、第 604662號、第 627752 號、第0267159號，以及第0292435號。 植物轉形之其他方法包括微喷射（microprojectile)-媒 介轉形，其中DNA係攜帶於微噴射之表面。該表現載體係 引入植物組織中，使用基因槍(biolistic)裝置，其可加速微 噴射粒子至足以穿透植物細胞壁與細胞膜之速度。參見， 例如：Sanford等人，Part· Sci· Technol. 5:27 (1987)，Sanford， J. C., Trends Biotech. 6:299 (1988), Sanford, J. C., Physiol. 88 201038734Such plants of the organism, or plants whose plants have been genetically engineered to produce these compounds/reagents. Thus, any plant species or plant cell can be selected. Examples of plant and plant cells, and plants grown or derived therefrom include, but are not limited to, plants and plant cells (turnips) obtainable from canola; rapeseed cultivars NQC02CNX12 (ATCC PTA-6011), NQC02CNX21 (ATCC PTA- 6644), and NQC02CNX25 (ATCC PTA-6012), as well as cultivars, breeding cultivars, and plant parts derived from rapeseed cultivars NQC02CNX12, NQC02CNX21, and NQC02CNX25 (see, respectively, US Patent No. 7,355,100, No. 7,456,340 'and 7, 348, 473); soybean (soya); rapeseed (brassica species), linseed/linen (linseed (Z/A7MW like / plus / heart / condition)); (Malay (Zea mfljAs)); safflower (red flower (Cari/zamw·? //«cior/wi)), cumin (to 曰Cai (good (10) 仙like)); 于草(于草{Nicotiana tabacum) ) 'Arabidopsis thaliana, Brazil chestnut (Brazilian chestnut CSei/w/eiiz'a ejcce/)); Castor bean (COwwMmi); coconut (cocon (Coc; Hu Wei (Hu Tuo) Cor/a«(irww ίαί/vwm)), cotton (cotton species; flowers (Arachis hypogaea); Jojoba (Himondwood boxwood c/nVzewA)); mustard (Brassica species and white mustard (57 secret alpha)); oil coconut (oil coconut (five / (10) ·? gw —m)); olives (column (〇/如 ewrpaea), rice (rice saiz.va)); pumpkin (pumpkin wax/wa), barley (barley (//ori/eww vw/g^re) ); wheat (wheat (TVaei/cww iiesi/vwm)), and duckweed (duckweed species • s/?)). Plant varieties from these and other plants can be produced, selected by 84 201038734, or optimized for the desired characteristics, such as below or associated with, but not limited to: seed yield, lodging resistance Sex, feathering, disease resistance or tolerance, maturity, late season plant integrity, plant height, cracking resistance, easy plant transformation, oil content, or oil form. Plant varieties can be selected by plant breeding, for example, breeding of purebred species, periodic screening breeding, xenogeneic mating and backcross breeding, and, for example, methods for marker assisted breeding and cultivation. See, for example, U.S. Patent No. 7,348,473. Animal cells include any isolated animal cells. The present invention is directed to a host cell that exhibits one or more nucleic acid molecules or recombinant nucleic acid molecules of the invention, including vectors. The present invention is directed to a method of making a recombinant host cell comprising introducing a recombinant vector into a host cell. The host cell can be genetically engineered (transduced or transfected or transfected) with a vector of the invention, which can be, for example, a selection vector or an expression vector. The vector may be, for example, in the form of a plastid, a viral particle, a bacteriophage or the like. The vector contains a polynucleotide sequence as described herein, and a suitable promoter or control sequence which can be used to transform a suitable host to permit expression of the polypeptide encoded by the polynucleotide. Genetic modification of the host cell can also include better or optimal gene optimization for host codon usage. The engineered host cell can be cultured in a conventional nutrient medium suitable for activating a promoter, screening a transformant, or amplifying a gene of the present invention. Culture conditions, such as temperature, pH, and the like, 85 201038734 are such culture conditions that were previously used for screening for host cell screening, and are apparent to those skilled in the art. In some embodiments, the invention is directed to genetically modifying a plant or part of a plant to express a PUFA synthase system as described herein, comprising at least a core PUFA synthase enzyme complex. "part of a plant" or, "piant part" as defined herein includes any part of a plant, such as, but not limited to, seeds (mature and immature), oil, Pollen, embryos, flowers, fruits, young shoots, leaves, roots, trunks, explants, etc. In some embodiments, the genetically modified plant or part of a plant produces one or more PUFAs 'eg, EPA, DHA, DPA (n-3 or n-6), ARA, GLA, SDA, other PUFAs , and combinations of them. It is not known that plants endogenously contain a PUFA synthase system; thus, the PUFA synthase system of the present invention can be used to engineer plants having unique fatty acid production capabilities. In a further embodiment, the plant or part of the plant is further genetically modified to express at least one PUFA synthase accessory protein (e.g., PPTase). In some embodiments, the plant is an oil seed plant, wherein the oil seed, and/or the oil in the oil seed, comprises a PUFA made by a PUFA synthase system. In some embodiments, the genetically modified plant, part of the plant, oil seed, and/or oil in the oil seed contains a detectable amount of at least one PUFA that is the PUFA synthase system product. In further embodiments, the oils in the plants, plant parts, oil seeds', and/or oil seeds may be substantially free of intermediates or by-products that are not the primary PUFA of the introduced PUFA synthase system. The product 'and which is not produced naturally by the endogenous FAS system in wild-type plants 86 201038734. Although wild-type plants produce short- or medium-chain PUFAs via the FAS system, such as 18-carbon PUFAs, new or additional puFAs may be found in plants, plant parts, oil seeds, and/or oil seeds. The production line in the oil is the result of genetic modification of the PUFA synthase system described herein. Genetic modification of plants can be achieved using typical strain development and/or molecular genetic techniques. See, U.S. Application Publication No. 2/7/244192. Methods for producing genetically transgenic plants are known to those skilled in the art, and a recombinant nucleic acid molecule encoding the desired amino acid sequence is incorporated into the genome of the plant. For example, a viral vector can be used to produce a genetically transgenic plant, for example, by transformation of a plant with a single cotyledon of a viral vector, which uses U.S. Patent No. 5,569,597; 5,589,367; and 5,316,931 The method described in the number. Methods for genetic engineering to modify or modify plants by transformation are also well known in the art, including biological and physical transformation processes. See, for example, 'BL Miki et al., Procedures for Introducing Foreign DNA into Plants, in METHODS IN PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY 67-88 (Glick, BR and Thompson, JE eds., CRC Press, Inc., Boca Raton, 1993) . In addition, vectors and in vitro culture methods for plant cell or tissue transformation and plant regeneration are available. See, for example, Μ·Y. Gruber et al., Vectors for Plant Transformation, in METHODS IN PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY 89-119 (Glick, B. R. and Thompson, J. E. eds., CRC Press, Inc., Boca Raton, 1993). 87 201038734 The widely used method for introducing a performance vector into plants is based on the natural transformation system of Agrobacterium. See, for example, Horsch et al., Science 227: 1229 (1985) and U.S. Patent No. 6, 〇 51, 757. A. tumefaciens and A. rhizogenes are plant pathological soil bacteria which can transform plant cells into genes. The Ti and Ri system of Agrobacterium tumefaciens and Agrobacterium rhizogenes each carry a corresponding gene that transforms the plant gene. See, for example, Kado, C. I., Crit. Rev. Plant. Sci. 10:1 (1991). Descriptions of methods for gene transfer of Agrobacterium vector systems and Agrobacterium vectors are provided in several literatures, including Gruber et al., supra; Miki et al., supra; Moloney et al., Plant Cell Reports 8: 238 (1989); U.S. Patent No. 5, 177, 010; No. 5, 149, 645; No. 5, 463, 174; No. 4, 762, 785; No. 5, 004, 803; and No. 5, 159, 135; and European Patent Application No. 0316624, No. 120516, No. 159418, No. 176112, No. 116718, No. 290799, No. 320500, No. 604662, No. 627752, No. 0267159, and No. 0 292 535. Other methods of plant transformation include microprojectile-mediated transformation in which the DNA is carried on the surface of the microprojection. The expression vector is introduced into plant tissue using a biolistic device that accelerates the microprojection of particles to a rate sufficient to penetrate the plant cell wall and the cell membrane. See, for example, Sanford et al., Part Sci·Technol. 5:27 (1987), Sanford, J. C., Trends Biotech. 6:299 (1988), Sanford, J. C., Physiol. 88 201038734

Plant 79:206 (1990)，Klein 等人，Biotechnology 10:268 (1992) ’以及美國專利案第5,015,580號與第5,322,783號。 加速塗覆於微粒子上的遺傳物質指向細胞内的技術亦說 明’例如’於美國專利案第4,945,050號與第5，141,141號之 中。物理性傳送DNA至植物中之另一種方法為將標的細胞 超音波震覆。參見，例如，Zhang等人，Bio/Technology 9:996 (1的1)。任擇地，脂質體或球狀體(spher〇piast)融合已用於 將表現載體引入至植物中。參見，例如，Deshayes等人， EMBO J·，4:2731 (1985) ’ Christou等人，Proc Natl. Acad. Sci. USA 84:3962 (1987)。亦已經報導使用CaCl2沈澱、DNA注 射、聚乙稀醇或聚-L-鳥胺酸(ornithine)來將DNA直接攝入 於原生質體(protoplast)中。參見，例如，Hain等人，Mol. Gen_ Genet· 199:161 (1985)與Draper等人，Plant Cell Physiol. 23:451 (1982)。原生質體以及全細胞與組織之電穿孔法也 已經被說明。參見，例如：Donn等人，in Abstracts of Vllth International Congress on Plant Cell and Tissue Culture IAPTC，A2-38, p. 53 (1990) ; D，Halluin等人，Plant Cell 4:1495-1505 (1992); Spencer等人，Plant Mol. Biol. 24:51-61 (1994);國際申請公開案WO 87/06614、W0 92/09696，和 W0 93/21335 ;以及美國專利案第5,472,869號與第5,384,253 號。其他的轉形技術包括晶鬚技術，參見，例如，美國專 利案第5,302,523號與第5,464,765號。 葉綠體或色素體也可以直接被轉形。確切而言，重組 型植物可經製造，其中僅葉綠體或色素體DNA已經用以上 89 201038734 說明的該等核酸分子及該等重組型核酸分子的任一者以及 其等之組合予以修飾。於葉綠體與色素體内作用的啟動子 係熟悉此藝者知道的。參見，例如，Hanley-Bowden等人， Trends in Bi〇chemical Sciences n:67-70 (1987)。用於獲得 含有已插入異源性dna之葉綠體的細胞之方法與組成物已 描述於’例如，美國專利案第5,693,507號與第5,451，513號 之中。 也可以使用提供有效的轉形之任何其他的方法。 適合使用於植物轉形的載體係熟悉此藝者知道的。參 見’例如：美國專利案第6,495,738號；第7,271,315號；第 7, 34M73號；第7,355,1〇〇號；第7,456,340號；以及其中揭 示的文獻。 表現載體能包括操作地連結至一調節要素（舉例而 言，一啟動子）之至少一個遺傳標諸，其允許含有該標德之 經轉形的細胞藉由負向選擇予以回收，亦即，抑制含有該 可選擇標誌基因的細胞之生長，或是藉由正向選擇予以回 收，亦即，4選由該遺傳標誌編碼的產物。許多普遍使用 之植物轉形的可選擇標誌基因係轉形技藝熟知的，以及包 括，舉例而言，編碼代謝上解毒一選擇性化學劑（其可以是 一抗生素或一除草劑)之酵素的基因，或是編碼一改變的標 的（對抑制子不敏感的）之基因。植物轉形中適合使用的巧·選 擇標誌包括，但不限於：轉位子Tn5的胺基醣苷磷酸轉移酶 基因（Aph II) ’其編碼抗生素康黴素、新黴素 (neomycin) ’和G418的抗性，以及編碼以下之抗性或是耐 90 201038734 受性的該等基因：草甘膦、潮黴素、甲氨蝶呤、膦基丁胺 酸(phosphinothricine)(畢拉草（bial〇ph〇s))、二氮雜戊烯類 (imidazolinones)、磺醯脲和三唑嘧啶除草劑，例如：氣磺 隆(chlorsulfuron)、溴苯腈(br〇rnoxynii)、茅草枯（dalapon)， 以及類似物。一種普遍使用之植物轉形的可選擇標誌基因 係於植物調控訊息的控制之下的新黴素磷酸轉移酶II(nptII) 基因’其賦予康黴素的抗性。參見，例如，Fraley等人-Proc· ΟPlant 79: 206 (1990), Klein et al, Biotechnology 10: 268 (1992) and U.S. Patent Nos. 5,015,580 and 5,322,783. The technique of accelerating the application of the genetic material on the microparticles to the cells is also described, for example, in U.S. Patent Nos. 4,945,050 and 5,141,141. Another method of physically transferring DNA into plants is to ultrasonically shock the target cells. See, for example, Zhang et al, Bio/Technology 9:996 (1 of 1). Optionally, liposomal or spher〇piast fusions have been used to introduce expression vectors into plants. See, for example, Deshayes et al, EMBO J., 4:2731 (1985) ‘Christou et al, Proc Natl. Acad. Sci. USA 84:3962 (1987). It has also been reported that CaCl2 precipitation, DNA injection, polyethylene glycol or poly-L-ornithine is used to directly ingest DNA into protoplast. See, for example, Hain et al, Mol. Gen_Genet. 199:161 (1985) and Draper et al, Plant Cell Physiol. 23:451 (1982). Protoplasts as well as electroporation of whole cells and tissues have also been described. See, for example, Donn et al, in Abstracts of Vllth International Congress on Plant Cell and Tissue Culture IA PTC, A2-38, p. 53 (1990); D, Halluin et al, Plant Cell 4: 1495-1505 (1992); Spencer et al., Plant Mol. Biol. 24: 51-61 (1994); International Application Publication Nos. WO 87/06614, WO 92/09696, and WO 93/21335; and U.S. Patent Nos. 5,472,869 and 5,384,253. Other morphing techniques include whisker technology, see, for example, U.S. Patent Nos. 5,302,523 and 5,464,765. The chloroplast or pigment body can also be directly transformed. Specifically, recombinant plants can be made wherein only chloroplast or chromoplast DNA has been modified with any of the nucleic acid molecules described above and the recombinant nucleic acid molecules described above and combinations thereof. Promoters that act in the chloroplast and pigment bodies are known to those skilled in the art. See, for example, Hanley-Bowden et al, Trends in Bi〇chemical Sciences n: 67-70 (1987). The method and composition for obtaining a cell containing a chloroplast into which a heterologous DNA has been inserted is described in, for example, U.S. Patent Nos. 5,693,507 and 5,451,513. Any other method that provides an effective transformation can also be used. Carriers suitable for use in plant transformation are well known to those skilled in the art. See, for example, U.S. Patent No. 6,495,738; U.S. Patent No. 7,271,315; No. 7,34, M. No. 7, 355, No. 7, No. 7, 456, 340; A performance vector can include at least one genetic marker operatively linked to a regulatory element (eg, a promoter) that allows transformed cells containing the marker to be recovered by negative selection, ie, The growth of cells containing the selectable marker gene is inhibited or recovered by positive selection, i.e., the product encoded by the genetic marker is selected. Many commonly used plant-transformable selectable marker genes are well known in the art of transformation, and include, for example, genes encoding enzymes that are metabolically detoxifying a selective chemical (which may be an antibiotic or a herbicide). Or a gene encoding a altered target (insensitive to the repressor). Suitable selection markers for plant transformation include, but are not limited to, the aminoglycoside phosphotransferase gene (Aph II) of the transposon Tn5, which encodes the antibiotics such as oxytetracycline, neomycin, and G418. Resistance, and the genes encoding the following resistance or resistance to 90 201038734: glyphosate, hygromycin, methotrexate, phosphinothricine (bial〇ric) 〇s)), imidazolinones, sulfonamides and triazolopyrimidine herbicides, for example: chlorsulfuron, br〇rnoxynii, dalapon, and analog. A commonly used plant-transformable selectable marker gene is the neomycin phosphotransferase II (nptII) gene, which is under the control of plant regulatory messages, which confers resistance to benzimycin. See, for example, Fraley et al. - Proc· Ο

Natl. Acad. Sci. U.S-A. 80: 4803 (1983)。另一個普遍使用之 可選擇標誌基因係潮黴素磷酸轉移酶基因，其賦予抗生素 潮徽素的抗性。參見，例如，VandenElzen等人，Plant Mol. Biol. 5:299 (1985)。細菌來源之額外的賦予抗生素抗性的可 選擇標諸基因包括健他徽素乙酸轉移酶(gentamycin acetyl transferase)、鏈黴素磷酸轉移酶、胺基醣苷_3，_腺苷酸轉移 酶，以及博歐黴素(bleomycin)抗性的決定位。參見，例如： Hayford等人，Plant Physiol. 86:1216 (1988)，Jones等人， Mol. Gen. Genet. 210: 86 (1987)，Svab等人，Plant Mol· Biol. 14:197 (1990) ’ Hille等人，Plant Mol_ Biol· 7:171 (1986)。 其他的可選擇標誌基因賦予除草劑的抗性，例如：草甘膦、 草丁膦(glufosinate)，或是溴苯腈。參見，例如，Comai等 人，Nature 317:741-744 (1985)，Gordon-Kamm等人，Plant Cell 2:603-618 (1990)以及Stalker等人，Science 242:419-423 (1988)。用於植物轉形之其他可選擇標誌基因為非細菌來源 的。此等基因包括，舉例而言，小鼠二氫葉酸還原酶、植 物的5-烯醇丙酮酸莽草酸-3-磷酸合成酶 91 201038734 (5-enolpyruvylshikimate-3-phosphate synthase)和植物乙醢 乳酸合成酶。參見，例如：Eichholtz等人，Somatic Cell Mol. Genet. 13:67 (1987)，Shah等人，Science 233:478 (1986)， Charest等人，Plant Cell Rep. 8:643 (1990)。 一報導基因能與一可選擇標誌一起使用或在沒有可選 擇標誌的情況下使用。報導基因係典型地不存在於接收者 有機體或組織内的基因，以及典型地編碼導致一些表現型 改變或是酵素的性質之蛋白質。參見，例如，K. Weising 等人，Ann· Rev. Genetics 22: 421 (1988)。報導基因包括， 但不限於：/3 —尿苷酸酶(GUS)、/3 —半乳糖苷酶、氣黴素 乙醯轉移酶、綠螢光蛋白質，以及螢光素酶基因。參見， 例如：Jefferson，R. A·，Plant Mol. Biol. Rep. 5:387 (1987)， Teeri等人，EMBO J. 8:343 (1989)，Koncz等人，Proc. Natl. Acad· Sci U.S.A. 84:131 (1987)，DeBlock等人，EMBO J· 3:1681 (1984)，以及Chalfie等人，Science 263:802 (1994)。 在基因已經引入至接收者細胞内之後的適合的時間可以執 行偵測報導基因表現之分析。一此分析必需使用編碼大腸 桿菌之uida位址的/3—尿苷酸酶(GUS)基因，如Jefferson等 人，Biochem· Soc· Trans. 15: 17-19 (1987)之中說明的。 來自各種各樣的來源之啟動子調節要素能於植物細胞 内有效地使用以表現外來的基因。舉例而言，能使用細菌 來源的啟動子調節要素，例如：章魚肉鹼合成酶啟動子、 胭脂鹼(nopaline)合成酶啟動子、甘露鹼(mannopine)合成酶 啟動子’以及病毒來源的啟動子，例如：花椰菜斑紋病毒 92 201038734 (35S和19S)、35T(其係經再改造的35S啟動子，參見國際申 請公開案WO 97/13402)。植物啟動子調節要素包括但不限 於：核酮酸-1，6-雙磷酸(RUBP)羧酶小次單元(ssu)、大豆 伴球蛋白（beta-conglycinin)啟動子、菜豆蛋白啟動子、 ADH啟動子、熱休克啟動子，以及組織專一性啟動子。基 質結合區域' 支架結合區域、内含子、增強子，以及聚腺 苷酸化序列也可以用來改良轉錄效率或是DNA的結合。此 專元素flb被包括以得到經轉形的DNA植物中之最佳的效 能。典型的元素包括，但不限於：Adh-内含子1、Adh_内含 子6、為蓿斑紋病毒外殼蛋白引導序列、玉蜀黍條斑病毒外 殼蛋白引導序列，以及熟悉此藝者可得的其餘物。構成性 啟動子調節要素也可以用來指示連續的基因表現。構成性 啟動子包括，但不限於：來自植物病毒的啟動子，例如來 自 CaMV 之 35S啟動子（Odell等人，Nature 313:810-812 (1985)) ’以及來自以下此等基因之啟動子，如：米的肌動Natl. Acad. Sci. U.S-A. 80: 4803 (1983). Another commonly used selectable marker gene is the hygromycin phosphotransferase gene, which confers resistance to the antibiotic tibia. See, for example, VandenElzen et al, Plant Mol. Biol. 5:299 (1985). Additional selectable genes that confer antibiotic resistance from bacterial sources include gentamycin acetyl transferase, streptomycin phosphotransferase, aglycone-3, adenylate transferase, and The determining position of bleomycin resistance. See, for example: Hayford et al, Plant Physiol. 86: 1216 (1988), Jones et al, Mol. Gen. Genet. 210: 86 (1987), Svab et al, Plant Mol. Biol. 14: 197 (1990) ' Hille et al., Plant Mol_ Biol. 7:171 (1986). Other selectable marker genes confer resistance to the herbicide, such as glyphosate, glufosinate, or bromoxynil. See, for example, Comai et al, Nature 317: 741-744 (1985), Gordon-Kamm et al, Plant Cell 2: 603-618 (1990) and Stalker et al, Science 242: 419-423 (1988). Other selectable marker genes for plant transformation are of non-bacterial origin. Such genes include, for example, mouse dihydrofolate reductase, plant 5-enolpyruvylshikimate-3-phosphate synthase 91 201038734 (5-enolpyruvylshikimate-3-phosphate synthase) and plant acetaminophen Synthetic enzyme. See, for example, Eichholtz et al, Somatic Cell Mol. Genet. 13:67 (1987), Shah et al, Science 233:478 (1986), Charest et al, Plant Cell Rep. 8:643 (1990). A reporter gene can be used with a selectable marker or without an optional marker. Reporter genes are typically genes that are not found in the recipient organism or tissue, as well as proteins that typically encode properties that result in some phenotypic changes or enzymes. See, for example, K. Weising et al., Ann. Rev. Genetics 22: 421 (1988). Reporter genes include, but are not limited to, /3 - uridine betainase (GUS), /3 - galactosidase, oxytetracycline acetyltransferase, green fluorescent protein, and luciferase gene. See, for example, Jefferson, R. A., Plant Mol. Biol. Rep. 5:387 (1987), Teeri et al, EMBO J. 8:343 (1989), Koncz et al, Proc. Natl. Acad·Sci USA 84: 131 (1987), DeBlock et al., EMBO J. 3: 1681 (1984), and Chalfie et al., Science 263: 802 (1994). The analysis of the reporter gene expression can be performed at a suitable time after the gene has been introduced into the recipient cell. For this analysis, it is necessary to use the /3-uridinease (GUS) gene encoding the uida site of E. coli, as described in Jefferson et al., Biochem Soc. Trans. 15: 17-19 (1987). Promoter regulatory elements from a variety of sources can be effectively used in plant cells to express foreign genes. For example, bacterial-derived promoter regulatory elements can be used, such as: octopine carnitine synthase promoter, nopaline synthase promoter, mannopine synthase promoter', and viral-derived promoters. For example: Cauliflower Streak Virus 92 201038734 (35S and 19S), 35T (which is a remodeled 35S promoter, see International Application Publication WO 97/13402). Plant promoter regulatory elements include, but are not limited to, ribulonic acid-1,6-diphosphate (RUBP) carboxylase small subunit (ssu), beta-conglycinin promoter, phaseolin promoter, ADH Promoters, heat shock promoters, and tissue-specific promoters. The matrix binding region's scaffold binding region, intron, enhancer, and polyadenylation sequences can also be used to improve transcriptional efficiency or DNA binding. This elemental element flb is included to give the best effect in transformed DNA plants. Typical elements include, but are not limited to, Adh-intron 1, Adh_intron 6, zebra virus coat protein leader sequence, maize leaf spot virus coat protein leader sequence, and others available to those skilled in the art. Things. Constitutive promoter regulatory elements can also be used to indicate continuous gene expression. Constitutive promoters include, but are not limited to, promoters derived from plant viruses, such as the 35S promoter from CaMV (Odell et al, Nature 313:810-812 (1985)) and promoters from these genes, Such as: muscle movement

蛋白（McElroy 等人，Plant Cell 2:163-171 (1990))、泛蛋白 (Christensen等人，Plant Mol. Biol. 12:619-632 (1989)及 Christensen等人，Plant Mol_ Biol. 18:675-689 (1992))、 pEMU(Last等人，Theor. Appl_ Genet· 81:581-588 (1991))、 MAS(Velten等人，EMBO J. 3:2723-2730 (1984))、玉蜀黍H3 組織蛋白（Lepetit等人，Mol· Gen. Genetics 231:276-285 (1992)和 Atanassova 等人，Plant Journal 2(3): 291-300 (1992))，以及ALS啟動子、Xbal/Ncol片段5·至西洋油菜 (Brassica napus)ALS3結構基因（或相似於Xbal/Ncol片段之 93 201038734 核苷酸序列Η國際申請公開案w〇 96/30530)。組織專一性 啟動子5周節要素也可以用於特定的細胞類或是組織類型中 的基因表現’例如：葉子或種子(例如：玉米蛋白（zein)、油 體膜蛋白（0丨eosin)、油菜籽蛋白（napin)、ACp、球蛋白，以 員U物）。組織專一性或組織偏好的啟動子包括，但不限 於.根偏好的啟動子，例如：來自菜豆蛋白基因（Murai等 人 Science 23:476-482 (1983)與Sengupta-Gopalan等人， Proc. Natl. Acad· Sci_ U.S.A· 82:3320-3324 (1985));葉子專 一性和光誘導性啟動子，例如：來自cab或是核酮糖雙磷酸 竣化酶(rubisco) (Simpson等人，EMBO J. 4(11):2723-2729 (1985)與Timko等人 ’ Nature 318:579-582 (1985));花藥專 一性啟動子’例如：來自LAT52(Twell等人，Mol. Gen. Genetics 217:240-245 (1989));花粉專一性啟動子，例如： 來自 Zml3(Guerrero等人，Mol. Gen. Genetics 244:161-168 (1993));或是小孢子-偏好的啟動子，例如：來自apg (Twell 等人，Sex. Plant Reprod· 6:217-224 (1993))。啟動子調節要 素也可以在植物發展的某些階段的期間，以及植物組織和 器官為活性的，包括，但不限於：花粉專一性、胚芽專一 性、玉米穗黃專一性、棉花纖維專一性、根專一性’以及 種子内胚乳專一性啟動子調節要素。能使用的一誘導性啟 動子調節要素，其負責對專一性的訊號反應之基因的表 現，例如：生理刺激(熱休克基因）；光(RUBP羧酶）；激素 (Em);代謝物；化學品；以及壓力。誘導性啟動子包括’ 但不限於：來自對銅反應之ACEI系統的一啟動子（Mett等 94 201038734 人，PNAS 90:4567-4571 (1993));來自玉蜀黍之In2基因， 其係對反應苯磺醯胺除草劑安全劑(Hershey等人，Mol. Gen Genetics 227:229-237 (1991)和 Gatz 等人，m〇1. Gen. Genetics 243:32-38 (1994))，來自 TnlO之Tet 抑制子(Gatz等 人，Mol_ Gen. Genetics 227:229-237 (1991));以及來自類固 醇激素基因，其之轉錄的活性係由醣皮質類固醇激素所誘 導（Schena 等人，Proc. Natl· Acad. Sci· U.S.A. 88:0421 (1991) 〇 ο 訊息序列也可以用來指示一多肽往一細胞内胞器或次 細胞隔室或是分泌至質外體。參見，例如：Becker等人， Plant Mol. Biol. 20:49 (1992) ’ Knox, C.等人，Plant Mol. Biol. 9:3-17 (1987)，Lerner 等人，Plant Physiol. 91:124-129 (1989) ’ Fontes等人，Plant Cell 3:483-496 (1991)，Matsuoka 等人，Proc· Natl. Acad. Sci. 88:834 (1991)，Gould等人，J. Cell. Biol· 108:1657 (1989)，Creissen等人，Plant J. 2:129 (1991)，Kalderon，等人，Cell 39:499-509 (1984)，以及Steifel 等人，Plant Cell 2:785-793 (199〇)。此等尋標序列提供待轉 移之所欲表現的蛋白質至其最有效地作用之細胞結構内， 或是至對於所欲的表現型功能為必須的細胞加工集中之細 胞的區域。 於一些具體例中，訊息序列被用來指示本發明的蛋白 質向一次細胞隔室，舉例而言，至色素體或葉綠體。基因 產物，包括異源性基因產物，能藉由將基因產物融合至一 訊息序列而尋標至色素體或葉綠體，該訊息序列可在葉綠 95 201038734 體輸入時被切割’而產生成熟蛋白質。參見，例如，c〇mai 等人 ’ J. Biol. Chem. 263: 15104-15109 (1988)與van den Broeck等人，Nature 313: 358-363 (1985)。編碼合適的訊息 序列之DNA可以自編碼RUBISCO蛋白質、CAB蛋白質、 EPSP合成酶酵素、GS2蛋白質的cDNA單離，或是自任何天 然存在的葉綠體尋標的蛋白質單離，該葉綠體尋標的蛋白 質含有會指示該尋標蛋白質向葉綠體的一訊息序列（亦稱 為葉綠體轉運肽(CTP))。此等葉綠體尋標蛋白質係本技藝 所熟知的。該等葉綠體尋標蛋白質係合成為較大的前驅物 蛋白質，其含有一胺基端CTP，其指示該前驅物往葉綠體 輸入機構。CTPs—般係以葉綠體胞器内之特異性内蛋白酶 切割，因而自該前驅物釋放該尋標成熟蛋白質，包括活性 蛋白質（例如酵素），至葉綠體環境。編碼胜肽，該胜肽適用 於引導標的基因或基因產物至植物細胞之葉綠體或色素體 中，之序列的實例包括牽牛花EPSPS CTP、阿拉伯芬 〇4raMi/o/?^)EPSPSCTP2與内含子，以及熟悉此藝者知道 的其他序列。CTPs之特定的實例包括，但不限於：阿拉伯 芥核酮醣雙填酸鹽叛化酶小次單元 atslA轉移胜肽、阿拉伯茶(JrahWopA 轉運 肽’以及玉米（Zea maize)核酮醣雙磷酸鹽羧化酶小次單元 轉運肽。最佳化的轉運肽係揭示於，例如，Van den Broeck 等人’ Nature 313:358-:363 (1985)中。原核與真核訊息序列 係揭示於，例如，Michaelis等人，Ann. Rev. Microbiol. 36: 425 (1982)中。能使用於本發明之中的轉運肽之額外的實例 96 201038734 包括 Von Heijne等人，Plant Mol. Biol. Rep. 9:104-126 (1991) ; Mazur等人，Plant Physiol. 85: 1110 (1987) ; Vorst 等人，Gene 65: 59 (1988); Chen & Jagendorf, J. Biol. Chem. 268: 2363_2367 (1993)之中說明的葉綠體轉運肽；來自菸草 CA/VconVma 之 rbcS 基因之轉運肽（Poulsen 等人，Mol. Gen. Genet. 205: 193-200 (1986));以及自西洋 油菜（价osWca παρΜ·?)醯基-ACP硫酯酶衍生的轉運肽 ❹ （Loader等人，Plant Mol. Biol. 23: 769-778 (1993) ; Loader 等人，Plant Physiol. 110:336-336 (1995)。 本發明之經基因修飾的植物可以進一步修飾以刪除或 是不活化一内源的脂肪酸合成酶，以降低與外源的PUFA合 成酶系統之内源的競爭丙二酿基C〇A，以增加有機體中的 丙二醯基CoA之位準，以及其等之組合。參見，例如，美 國申請公開案第2007/0245431號。 經基因修飾的植物能於發酵培養基中予以培養或是於 Q 適合的培養基中生長，例如：土壤。高等植物之適合的生 長培養基包括任何植物之生長培養基，例如，但不限於： 土壤、沙子、任一其他可支撐根部生長之特定介質（如蛭石 (vermiculite)、珍珠岩(perlite)等，或是水栽培養，以及適合 的光線、水與營養補充物，其使高等植物之生長最佳化。 PUFAs能自經基因修飾的植物經由純化過程予以回收，其 自植物_萃取化合物。PUFAs能藉由收穫該植物而回收， 乂及藉由自 >>罐物收穫油而回收(例如，自油類種子）。該植 物也可以於其之自然狀態消耗或更進一步處理為可消耗產 97 201038734 品。於一些具體例中，本發明係針對經基因修飾的植物， 其中該植物生產由於基因修飾之至少一個PUFA，以及其中 植物中的總脂肪酸形態，或是該植物累積PUFAs的部分， 包含由於該植物的基因修飾所生產之可偵測量的PUFA。於 一些具體例中，該植物為油類種子植物。於一些具體例中， 該油類種子植物生產PUFAs於其之成熟種子中或含有 PUFAs於其之種子的油中。 也可以使用各種各樣的哺乳動物培養系統以表現重組 型蛋白質。表現載體會包含一複製起點、一適合的啟動子 與增強子’以及核糖體結合位址、聚腺苷酸化位址、剪接 供體與接受體位址、轉錄終止序列，和5’側邊未轉錄序列也 是必須的。 涉及異源性表現的方法 本發明係針對一種生產至少一個PUFA的方法，其包 含：於有效產生PUFA的條件下於一宿主細胞中表現puFA 合成酶系統，其中該PUFA合成酶系統包含本文中說明的該 等經單離的核酸分子及重組型核酸分子的任一者，以及其 等之組合，其中至少一個PUFA係被生產。於一些具體例 中，該至少一個PUFA包括DHA、EPA，或其等之一組合。 於一些具體例中，該宿主細胞係植物細胞、經單離的動物 細胞，或微生物細胞。於一些具體例中，該宿主細胞為破 囊壺菌。 本發明係針對一種生產富含DHA、EPA，或其等之— 組合的脂質的方法，其包含：於有效產生脂質的條件下於 98 201038734 一宿主細胞中表現PUFA合成酶基因，其中該PUFA合成酶 基因包含本文中說明的該等經單離的核酸分子及重組型核 酸分子的任一者以及其等之組合於該宿主細胞中，其中富 含DHA、EPA，或其等之一組合的脂質被生產。 本發明係針對一種自宿主細胞單離脂質的方法，其包 含於有效產生脂質的條件下於該宿主細胞中表現PUFA合 成酶基因，以及自該宿主細胞單離脂質，其中該宿主細胞 中的該PUFA合成酶系統包含本文中說明的該等經單離的 核酸分子及重組型核酸分子的任一者，以及其等之組合。 於一些具體例中，含有PUFAs之一或更多個脂質餾分 係自該等宿主細胞予以單離。於一些具體例中，自該宿主 細胞單離的一或更多個餾分包括總脂肪酸餾分、固醇酯餾 分、三酸甘油酯餾分、游離脂肪酸餾分、固醇餾分、雙甘 油餾分、磷脂質餾分，或其等之組合。於一些具體例中， PUFAs係自該等宿主細胞予以單離，其中該等PUFAs係富含 亞米加-3脂肪酸亞米加-6脂肪酸，或其等之組合，根據引入 至一宿主細胞中的該PUFA合成酶系統的組成。於一些具體 例中，該等PUFAs係富含DHA、EPA、DPA n-6、ARA，或 其等之組合，根據引入至一宿主細胞中的該PUFA合成酶系 統的組成。於一些具體例中，該等PUFAs係富含DHA、EPA， 或其等之一組合。於一些具體例中，自一宿主細胞單離的 PUFAs之PUFA形態包括高濃度的DHA和較低濃度的EPA、 ARA、DPAn-6，或其等之組合。於一些具體例中，自一宿 主細胞單離的PUFAs之PUFA形態包括高濃度的DHA與 99 201038734 EPA，和較低濃度的ARA、DPAn-6，或其等之組合。於一 些具體例中，自一宿主細胞單離的PUFAs之PUFA形態包括 高濃度的EPA和較低濃度的DHA、ARA、DPAn-6，或其等 之組合。 本發明係針對一種於具有p UFA合成酶活性的有機體 内取代不活化或被刪除的PUFA合成酶活性，引入新的 PUFA合成酶活性，或是增強現存的PUFA合成酶活性的方 法，其包含於有效表現PUFA合成酶活性的條件下於該有機 體中表現本文中說明的該等經單離的核酸分子及重組型核 酸分子的任一者及其等之組合。於一些具體例中，該核酸 分子包含本文中說明的一或更多個PiM/、，或是 PUFA合成酶多核苷酸序列，其係編碼一或更多個PUFA合 成酶領域。於一些具體例中，該等有機體的PUFA形態係藉 由引入本發明的一或更多個核酸分子予以改變。於一些具 體例中，該等經改變的PUFA形態包括亞米加-3脂肪酸的增 加以及亞米加-6脂肪酸的減少。於一些具體例中，該等經 改變的PUFA形態包括亞米加-6脂肪酸之增加以及亞米加-3 脂肪酸的減少。於一些具體例中，亞米加-3與亞米加-6脂肪 酸二者均為增加的。於一些具體例中，DHA的量為增加的， 而EPA、ARA、DPAn-6，或其等之組合的一或更多者的量 為維持或減少的。於一些具體例中’ EPA與DHA的量為增 加的，而ARA、DPAn-6，或其等之一組合的量為維持或減 少的。於一些具體例中’ EPA的量為增加的’而EPA、ARA、 DPA n-6，或其等之組合的一或更多者的量為維持或減少 100 201038734 的。於一些具體例中，該核酸分子包含PE43的多核苷酸序 列或其中的一或更多個領域。於一些具體例中，該核酸分 子包含PFJ3的多核苷酸序列或是其中的一或更多個領域’ 以及於該有機體中之亞米加-3脂肪酸的量是增加的而亞米 加-6脂肪酸的量為減少的。於一些具體例中，該核酸分子 包含ΡΛ42的多核苷酸序列或是其中的一或更多個領域，以 及於該有機體中之DHA的量是增加的而EPA的量為滅少 的。 本發明係針對在具有PUFA合成酶活性的有機體内增 加DHA、EPA，或其等之一組合的生產之方法，其包含於 有效產生DHA、EPA或其等之一組合的條件下於一有機體 中表現本文中說明的該等經單離的核酸分子及重組型核酸 分子的任一者之以及其等之組合，其中該PUFA合成酶活性 取代該有機體中之不活化或被刪除的活性，引入一新的活 性，或提高現存的活性，以及其中增加該有機體中的DHA、 EPA，或其等之一組合之生產。 本發明已經普遍地說明，可以藉由參考本文中提供的 實施例進一步的了解本發明。此等的實施例係僅為了闡釋 的目的而提供且不意欲為限制性的。 實施例1 設計KS與DH PUFA合成酶領域的簡併性引子俾以單 離來自以ATCC寄存編號PTA-9695寄存的經單離的微生 物，亦知道為裂殖壺菌種ATCCPTA-9695之對應序列。 裂殖壺菌種ATCC PTA-9695 PFJi的KS區域（亦即，含 101 201038734 有ks領域的區域)的簡併性引子係根據日本沙雷菌、裂殖壺 菌種ATCC 20888、金黃色破囊壺菌（ATCC 34304)，以及破 囊壺菌種23B ATCC 20892之公開的先前稱為orfA或 ORF 1)序列來設計： prDS173(順向）：GATCTACTGCAAGCGCGGNGGNTTYAT (序列辨識編號：62)，以及 prDS174(反向）：GGCGCAGGCGGCRTCNACNAC (序 列辨識編號：63)。 裂殖壺菌種ATCC PTA-9695 PFA3的DH區域(先前稱為 orfC或ORF 3)的簡併性引子係根據馬里納莫里特拉菌 (Μοηϊβ//α maWwa);裂殖壺菌種ATCC 20888 ;沙雷菌種 80尺02738;深處發光菌（/^〇/〇6(3<:化/-/7,〇!/^«办》2);以及破 囊壺菌種23B ATCC 20892之公開的序列來設計： JGM190(順向）：CAYTGGTAYTTYCCNTGYCAYTT (序 列辨識編號：64);以及 BLR242(反向）：CCNGGCATNACNGGRTC (序列辨識 編號：65)。 染色體DNA模板之PCR條件係如下：〇·2 μΜ dNTPs、 0.1 uM各引子、8% DMSO、200 ng 染色體DNA、2.5 U Herculase® II融合聚合酶（Stratagene)，以及IX Herculase® 緩衝液(Stratagene)配於50 μί總體積中。PCR流程包括以下 步驟：（1) 98°C歷時3分鐘；（2) 98°C歷時3〇秒；（3) 50°C歷 時30秒；（4) 72°C歷時2分鐘；（5)重複步驟2-4計40個循環； (6)72°C歷時5分鐘；以及(7)維持在6°C。 102 201038734 關於兩引子對，PCR使用來自裂殖壺菌種ATCC寄存編 號PTA-9695之染色體模板出產具有預期的大小之有區別的 DNA產物。各別的PCR產物係依據製造者之指示而被選殖 至該載體pJETl.2/鈍的（Fermentas)之内，以及使用所提供的 標準引子來決定插入序列。 該等從PCR產物得到的DNA序列係於標準的BLASTx 搜尋中與由NCBI GenBank可得的已知序列比較(BLASTx 參數：低複雜度過濾器開啟；矩陣：BLOSUM62 ;空格成 本；存在 11，擴展 l(Extenstionl)。Stephen F· Altschul, Thomas L. Madden, Alejandro A. Schaffer, Jinghui Zhang, Zheng Zhang, Webb Miller, and David J. Lipman (1997), "Gapped BLAST and PSI-BLAST : a new generation of protein database search programs", Nucleic Acids Res. 25:3389-3402)。 於胺基酸的位準，與含有來自裂殖壺菌種ATCC PTA-9695的KS片段之選殖的DNA所衍生之演繹的胺基酸 序列具有最高位準的同源性之序列為：裂殖壺菌種ATCC 20888 ”多不飽和脂肪酸合成酶次單元A"(同一性=87% ;陽 性=92%);沙雷菌（572ewa«e//a owezWe似z\y) MR-1 "多領域石-酮基醯基合成酶”（同一性=49% ;陽性=64%);以及沙雷菌 種MR-4 ”冷-嗣基醯基合成酶"（同一性=49%;陽性=64%)。Protein (McElroy et al, Plant Cell 2: 163-171 (1990)), ubiquitin (Christensen et al, Plant Mol. Biol. 12: 619-632 (1989) and Christensen et al, Plant Mol_ Biol. 18: 675 -689 (1992)), pEMU (Last et al., Theor. Appl_Genet 81:581-588 (1991)), MAS (Velten et al., EMBO J. 3: 2723-2730 (1984)), Jade H3 organization Protein (Lepetit et al, Mol. Gen. Genetics 231:276-285 (1992) and Atanassova et al, Plant Journal 2(3): 291-300 (1992)), and the ALS promoter, Xbal/Ncol fragment 5· To the Brassica napus ALS3 structural gene (or similar to the Xbal/Ncol fragment 93 201038734 nucleotide sequence Η International Application Publication No. 96/30530). Tissue-specific promoters 5-weekly elements can also be used for gene expression in specific cell types or tissue types 'eg leaves or seeds (eg zein, oil membrane protein (0 丨eosin), Rapeseed protein (napin), ACp, globulin, and U. Promoters that organize specificity or tissue preferences include, but are not limited to, root-preferred promoters, for example, from the Bean Protein Gene (Murai et al. Science 23: 476-482 (1983) and Sengupta-Gopalan et al., Proc. Natl Acad·Sci_ USA· 82:3320-3324 (1985)); Leaf-specific and light-inducible promoters, for example, from cab or ribulose-bisphosphonase (rubisco) (Simpson et al., EMBO J. 4(11): 2723-2729 (1985) and Timko et al. ' Nature 318: 579-582 (1985)); anther specific promoter 'for example: from LAT52 (Twell et al., Mol. Gen. Genetics 217: 240) -245 (1989)); pollen-specific promoters, for example: from Zml3 (Guerrero et al, Mol. Gen. Genetics 244:161-168 (1993)); or microspore-preferred promoters, eg from Apg (Twell et al., Sex. Plant Reprod. 6:217-224 (1993)). Promoter regulatory elements may also be active during certain stages of plant development, as well as plant tissues and organs, including, but not limited to, pollen specificity, germ specificity, maize ear specificity, cotton fiber specificity, Root specificity' and seed endosperm-specific promoter regulatory elements. An inducible promoter regulatory element that can be used to perform a gene response to a specific signal, such as: physiological stimulation (heat shock gene); light (RUBP carboxylase); hormone (Em); metabolite; chemistry Product; and pressure. Inducible promoters include, but are not limited to, a promoter from the ACEI system that responds to copper (Mett et al. 94 201038734, PNAS 90: 4567-4571 (1993)); the In2 gene from maize, which is a pair of reactive benzene Sulfamethine herbicide safeners (Hershey et al, Mol. Gen Genetics 227: 229-237 (1991) and Gatz et al, m〇1. Gen. Genetics 243: 32-38 (1994)), Tet from TnlO Inhibitors (Gatz et al., Mol_ Gen. Genetics 227: 229-237 (1991)); and from steroid hormone genes whose transcriptional activity is induced by glucocorticosteroids (Schena et al., Proc. Natl Acad) Sci· USA 88:0421 (1991) 〇ο The message sequence can also be used to indicate a polypeptide to an intracellular or subcellular compartment or to an apoplast. See, for example, Becker et al., Plant Mol. Biol. 20:49 (1992) 'Knox, C. et al., Plant Mol. Biol. 9:3-17 (1987), Lerner et al., Plant Physiol. 91:124-129 (1989) 'Fates et al. Human, Plant Cell 3:483-496 (1991), Matsuoka et al, Proc. Natl. Acad. Sci. 88:834 (1991), Gould et al, J. Cell. Biol. 8:1657 (1989), Creissen et al, Plant J. 2: 129 (1991), Kalderon, et al, Cell 39: 499-509 (1984), and Steifel et al, Plant Cell 2: 785-793 (199)寻) These search sequences provide the desired protein to be transferred into the cell structure in which it is most effective, or to the region of the cell in which the cell processing is necessary for the desired phenotypic function. In some embodiments, the message sequence is used to indicate that the protein of the invention is directed to a cell compartment, for example, to a chromosome or chloroplast. The gene product, including the heterologous gene product, can be fused to the gene product by The message sequence is searched for the chromoplast or chloroplast, and the message sequence can be cleaved upon input of chloroplast 95 201038734 to produce a mature protein. See, for example, c〇mai et al.' J. Biol. Chem. 263: 15104 -15109 (1988) and van den Broeck et al, Nature 313: 358-363 (1985). The DNA encoding the appropriate message sequence can be isolated from the cDNA encoding the RUBISCO protein, CAB protein, EPSP synthetase enzyme, GS2 protein, or isolated from any naturally occurring chloroplast-targeted protein, which indicates that the chloroplast-targeted protein contains The message sequence of the homing protein to the chloroplast (also known as the chloroplast transit peptide (CTP)). Such chloroplast-finding proteins are well known in the art. These chloroplast-finding proteins are synthesized as larger precursor proteins containing an amine-terminated CTP which indicates the precursor to the chloroplast input machinery. CTPs are typically cleaved by specific endoproteinases within the chloroplast organelles, thereby releasing the sought-after mature proteins, including active proteins (e.g., enzymes), from the precursor to the chloroplast environment. Encoding a peptide which is suitable for directing the target gene or gene product into the chloroplast or pigment body of the plant cell, examples of which include the morning glory EPSPS CTP, the Arabidopsis 4raMi/o/?^) EPSPSCTP2 and the intron And familiar with other sequences known to the artist. Specific examples of CTPs include, but are not limited to, Arabidopsis ribulose bisulphate dehydrogenase small subunit atslA transfer peptide, Arabic tea (JrahWopA transit peptide', and Zea maize ribulose bisphosphate Carboxylase small subunit transit peptides. Optimized transit peptide systems are disclosed, for example, in Van den Broeck et al., Nature 313:358-:363 (1985). Prokaryotic and eukaryotic message sequences are disclosed, for example, , Michaelis et al, Ann. Rev. Microbiol. 36: 425 (1982). Additional examples of transit peptides that can be used in the present invention 96 201038734 includes Von Heijne et al, Plant Mol. Biol. Rep. 9: 104-126 (1991); Mazur et al, Plant Physiol. 85: 1110 (1987); Vorst et al, Gene 65: 59 (1988); Chen & Jagendorf, J. Biol. Chem. 268: 2363_2367 (1993) The chloroplast transit peptide described therein; the transit peptide from the rbcS gene of tobacco CA/VconVma (Poulsen et al., Mol. Gen. Genet. 205: 193-200 (1986)); and from the western rape (price osWca παρΜ·?醯-ACP thioesterase-derived transit peptide ❹ (Loader et al, Plant Mol. Biol. 23: 769-778 (1993); Loader et al, Plant Physiol. 110:336-336 (1995). The genetically modified plants of the invention may be further modified to delete or not activate an endogenous fatty acid synthase to reduce Competing with the endogenous source of the exogenous PUFA synthetase system to increase the level of the propylenediamine-based CoA in the organism, and combinations thereof, see, for example, US Application Publication No. 2007/0245431. Genetically modified plants can be cultured in a fermentation medium or grown in a suitable medium such as soil. Suitable growth media for higher plants include growth media for any plant, such as, but not limited to : soil, sand, any other medium that supports root growth (such as vermiculite, perlite, etc., or hydroponics, and suitable light, water, and nutrient supplements that make higher plants Optimization of growth. PUFAs can be recovered from genetically modified plants via a purification process, which extracts compounds from plants. PUFAs can be recovered by harvesting the plant, and recovered by harvesting oil from >> cans (e.g., from oil seeds). The plant can also be consumed in its natural state or processed further as a consumable product 97 201038734. In some embodiments, the invention is directed to a genetically modified plant, wherein the plant produces at least one PUFA genetically modified, and wherein the total fatty acid form in the plant, or a portion of the plant that accumulates PUFAs, comprises Genetically modified to produce a detectable amount of PUFA. In some embodiments, the plant is an oil seed plant. In some embodiments, the oil seed plant produces PUFAs in mature seeds thereof or oils containing PUFAs in their seeds. A wide variety of mammalian culture systems can also be used to express recombinant proteins. The expression vector will contain an origin of replication, a suitable promoter and enhancer' and a ribosome binding site, a polyadenylation site, a splice donor and acceptor site, a transcription termination sequence, and a 5' flanking untranscribed Sequences are also required. Method of Involving Heterologous Expression The present invention is directed to a method of producing at least one PUFA comprising: expressing a puFA synthetase system in a host cell under conditions effective to produce PUFA, wherein the PUFA synthase system comprises the instructions herein Any of the isolated nucleic acid molecules and recombinant nucleic acid molecules, and combinations thereof, wherein at least one PUFA is produced. In some embodiments, the at least one PUFA comprises a combination of DHA, EPA, or the like. In some embodiments, the host cell is a plant cell, an isolated animal cell, or a microbial cell. In some embodiments, the host cell is a Thraustochytrium. The present invention is directed to a method for producing a lipid rich in DHA, EPA, or the like, comprising: expressing a PUFA synthase gene in a host cell in 98 201038734 under conditions effective to produce a lipid, wherein the PUFA synthesis The enzyme gene comprises any one of the isolated nucleic acid molecules and recombinant nucleic acid molecules described herein, and combinations thereof, in the host cell, wherein the lipid is enriched in a combination of DHA, EPA, or a combination thereof Being produced. The present invention is directed to a method of isolating a lipid from a host cell, comprising expressing a PUFA synthase gene in the host cell under conditions effective to produce a lipid, and isolating the lipid from the host cell, wherein the host cell The PUFA synthase system comprises any of the isolated nucleic acid molecules and recombinant nucleic acid molecules described herein, and combinations thereof. In some embodiments, one or more lipid fractions containing PUFAs are isolated from the host cells. In some embodiments, one or more fractions isolated from the host cell include a total fatty acid fraction, a sterol ester fraction, a triglyceride fraction, a free fatty acid fraction, a sterol fraction, a diglycerin fraction, a phospholipid fraction, , or a combination thereof. In some embodiments, the PUFAs are isolated from the host cells, wherein the PUFAs are rich in sub-male-3 fatty acid sub-plus-6 fatty acids, or combinations thereof, according to introduction into a host cell. The composition of the PUFA synthase system. In some embodiments, the PUFAs are enriched in DHA, EPA, DPA n-6, ARA, or a combination thereof, based on the composition of the PUFA synthase system introduced into a host cell. In some embodiments, the PUFAs are enriched in DHA, EPA, or a combination thereof. In some embodiments, the PUFA morphology of PUFAs isolated from a host cell comprises a high concentration of DHA and a lower concentration of EPA, ARA, DPAn-6, or combinations thereof. In some embodiments, the PUFA morphology of PUFAs isolated from primary host cells comprises a combination of high concentrations of DHA and 99 201038734 EPA, and lower concentrations of ARA, DPAn-6, or the like. In some embodiments, the PUFA morphology of PUFAs isolated from a host cell comprises a high concentration of EPA and a lower concentration of DHA, ARA, DPAn-6, or combinations thereof. The present invention is directed to a method for introducing a new PUFA synthase activity, or enhancing an existing PUFA synthase activity, by substituting an inactivated or deleted PUFA synthase activity in an organism having p UFA synthase activity, which is included in Any of the isolated nucleic acid molecules and recombinant nucleic acid molecules described herein, and combinations thereof, are expressed in the organism under conditions effective to express PUFA synthase activity. In some embodiments, the nucleic acid molecule comprises one or more PiM/, or PUFA synthase polynucleotide sequences described herein encoding one or more PUFA synthase domains. In some embodiments, the PUFA morphology of the organisms is altered by the introduction of one or more nucleic acid molecules of the invention. In some embodiments, the altered PUFA morphology includes an increase in the amiga-3 fatty acid and a decrease in the amiga-6 fatty acid. In some embodiments, the altered PUFA morphology comprises an increase in the amiga-6 fatty acid and a decrease in the amiga-3 fatty acid. In some embodiments, both Amiga-3 and Amiga-6 fatty acids are increased. In some embodiments, the amount of DHA is increased, while the amount of one or more of the combination of EPA, ARA, DPAn-6, or the like is maintained or reduced. In some embodiments, the amount of EPA and DHA is increased, while the amount of ARA, DPAn-6, or a combination thereof is maintained or reduced. In some embodiments, the amount of 'EPA is increased' and the amount of one or more of EPA, ARA, DPA n-6, or combinations thereof, is maintained or reduced by 100 201038734. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence of PE43 or one or more regions thereof. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence of PFJ3 or one or more of the domains 'and the amount of the amiga-3 fatty acid in the organism is increased and the amiga-6 The amount of fatty acids is reduced. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence of ΡΛ42 or one or more of the domains, and the amount of DHA in the organism is increased while the amount of EPA is reduced. The present invention is directed to a method of increasing the production of a combination of DHA, EPA, or the like in an organism having PUFA synthetase activity, which is contained in an organism under conditions effective to produce a combination of DHA, EPA, or the like. Characterizing any of the isolated nucleic acid molecules and recombinant nucleic acid molecules described herein, and combinations thereof, wherein the PUFA synthase activity replaces an activity that is not activated or deleted in the organism, introducing a The new activity, or the enhancement of the existing activity, and the production of a combination of one of DHA, EPA, or the like in the organism. The present invention has been generally described, and the present invention can be further understood by referring to the embodiments provided herein. The embodiments are provided for the purpose of illustration only and are not intended to be limiting. Example 1 Degenerate primers in the field of designing KS and DH PUFA synthase 单 to isolate the detached microorganisms deposited from ATCC accession number PTA-9695, also known as the corresponding sequence of Schizochytrium species ATCCPTA-9695 . The degenerate primer of the KS region of the Schizochytrium sp. ATCC PTA-9695 PFJi (ie, the region containing the ks field of 101 201038734) is based on Serratia marcescens, Schizochytrium sp. ATCC 20888, golden yellow sac The bacterium (ATCC 34304), and the previously known sequence of orfA or ORF 1) disclosed by Thraustochytrium sp. 23B ATCC 20892: prDS173 (forward): GATCTACTGCAAGCGCGGNGGNTTYAT (SEQ ID NO: 62), and prDS174 (reverse) To): GGCGCAGGCGGCRTCNACNAC (sequence identification number: 63). The degenerate primer of the DH region of the Schizochytrium strain ATCC PTA-9695 PFA3 (formerly known as orfC or ORF 3) is based on the Mori Moriella strain (Μοηϊβ//α maWwa); Schizochytrium ATCC 20888; Serratia species 80 feet 02738; deep luminescent bacteria (/^〇/〇6 (3<:化/-/7, 〇!/^«办) 2); and thraustochytrid strain 23B ATCC 20892 The disclosed sequence was designed to: JGM190 (forward): CAYTGGTAYTTYCCNTGYCAYTT (SEQ ID NO: 64); and BLR242 (reverse): CCNGGCATNACNGGRTC (SEQ ID NO: 65). The PCR conditions for the chromosomal DNA template are as follows: 〇·2 μΜ dNTPs, 0.1 uM primers, 8% DMSO, 200 ng chromosomal DNA, 2.5 U Herculase® II fusion polymerase (Stratagene), and IX Herculase® buffer (Stratagene) in a total volume of 50 μί. The PCR protocol includes the following Steps: (1) 98 ° C for 3 minutes; (2) 98 ° C for 3 seconds; (3) 50 ° C for 30 seconds; (4) 72 ° C for 2 minutes; (5) repeat steps 2 4 counts 40 cycles; (6) 72 ° C for 5 minutes; and (7) maintained at 6 ° C. 102 201038734 For the two primer pairs, PCR uses from the Schizochytrium strain ATCC The chromosome template of accession number PTA-9695 produces a differentiated DNA product of the expected size. The individual PCR products are selected to the vector pJETl.2/Fermentas according to the manufacturer's instructions. And using the provided standard primers to determine the insertion sequence. The DNA sequences obtained from the PCR products are compared to known sequences available from NCBI GenBank in a standard BLASTx search (BLASTx parameters: low complexity filter on; Matrix: BLOSUM62; space cost; presence 11, extension l (Extenstionl). Stephen F. Altschul, Thomas L. Madden, Alejandro A. Schaffer, Jinghui Zhang, Zheng Zhang, Webb Miller, and David J. Lipman (1997), &quot ;Gapped BLAST and PSI-BLAST : a new generation of protein database search programs", Nucleic Acids Res. 25:3389-3402). At the level of the amino acid, the sequence with the highest level of homology to the deduced amino acid sequence derived from the DNA of the KS fragment from Schizochytrium ATCC PTA-9695 is: Colony strain ATCC 20888 "polyunsaturated fatty acid synthase subunit A" (identity = 87%; positive = 92%); Serratia (572ewa «e / / a owezWe like z \ y) MR-1 &quot Multi-domain stone-ketothiol synthase" (identity = 49%; positive = 64%); and Serratia species MR-4 "cold-mercaptopurine synthetase" (identity = 49%) ; positive = 64%).

於胺基酸位準，與含有來自裂殖壺菌種ATCC PTA-9695之DH片段的選殖的DNA所衍生之演繹的胺基酸 序列具有最高位準的同源性之序列為：裂殖壺菌種ATCC 103 201038734 20888 "多不飽和脂肪酸合成酶次單元匚”（同一性=61%;陽 性=71%);沙雷菌（57^冰《狀//〇!/^<3/如《(3)人1'(1^ 700345 ”8-羥_基醯基-(醢基-載體-蛋白質）脫水酶FabA/FabZ”（同一性= 35% ;陽性=50%);以及沙雷菌 HAW-EB3，，亞米加-3多不飽和脂肪酸合成酶PfaC"(同一性= 34% ;陽性=50%)。 實施例2The amino acid sequence has the highest level of homology to the deduced amino acid sequence derived from the cloned DNA from the DH fragment of the Schizochytrium ATCC PTA-9695: fission The genus ATCC 103 201038734 20888 " polyunsaturated fatty acid synthase subunit 匚" (identity = 61%; positive = 71%); Serratia (57 ^ ice "//〇!/^<3 /eg, "(3) human 1' (1^ 700345" 8-hydroxy-ylindolyl-(mercapto-carrier-protein) dehydratase FabA/FabZ" (identity = 35%; positive = 50%); Serratia HAW-EB3, sub-male-3 polyunsaturated fatty acid synthase PfaC" (identity = 34%; positive = 50%).

PUFA合成酶基因係被鑑定來自裂殖壺菌種ATCC PTA-9695 。 基因體DNA係藉由標準的程序自微生物製備。參見， 例如，Sambrook J. and Russell D. 2001. Molecular cloning: A laboratory manual, 3rd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York。簡言之： (1)500 mL的細胞係自對數中期培養沉澱。再旋轉該等細 胞’以及全部微量的液體係用小孔吸管自該細胞沉澱丸移 除，（2)用 200pL溶解緩衝液(20mM Tris pH 8.0、125 pg/rnL 蛋白酶K、5〇mM NaCl、10mM EDTA pH 8.0、0.5% SDS) 將沉澱丸再懸浮；（3)細胞係在5〇〇C下溶解歷時i小時；（4 溶解的混合物係用吸量管移至鎖相凝膠(PLG -Eppendorf) 2mL·*# ; (5)添加等體積的p:c:l以及允許混合歷時丨5小時； (6)將官子以12k x g離心歷時5分鐘；（7)將水相自plg管内 的凝膠上予以移除且將等體積的氣仿添加至該水相，以及 予以混合歷時30分鐘；（8)將管子以14k離心歷時大概5分 鐘’（9)該頂層（水相）係用吸量管移離開氣仿，以及放置於 104 201038734 一新的管子内；（10)添加0 J體積的3M NaOAC以及予以混 合(倒轉數次）；（11)添加2倍體積的1〇〇% EtOH以及與此階 段形成的基因體DNA沉澱物混合(倒轉數次）；（丨2)該等管子 係於一微型離心管中在4t下以14k旋轉歷時大概15分鐘； (13)液體係與管子的底部剩餘的基因體dna被溫和地傾倒 掉；（14)用〇.5mL 70% EtOH清洗沉澱丸；（15)該等管子係於 一微型離心管中在4。(：下以14k旋轉歷時大概5分鐘；（16) 溫和地傾倒掉EtOH且乾燥基因體DNA沉澱丸；以及（17)適 當體積的ΗζΟ與RNase係被直接地添加至基因體DNA沉澱 丸。 該經單離的基因體DNA係用來產生由大片段（大概40 kB)所組成的重組庫（依據製造者之指示）於該黏接質體 pWEB-TNC™ (Epicentre)中。該黏接質體庫係藉由使用32P 放射性標記的探針之標準的群落雜交程序予以篩選 (Sambrook J. and Russell D. 2001. Molecular cloning: A laboratory manual, 3rd edition. Cold Spring HarborThe PUFA synthase gene line was identified from the Schizochytrium sp. ATCC PTA-9695. Genomic DNA is prepared from microorganisms by standard procedures. See, for example, Sambrook J. and Russell D. 2001. Molecular cloning: A laboratory manual, 3rd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York. Briefly: (1) A 500 mL cell line was cultured from a mid-log phase. Rotate the cells again and remove all traces of the liquid system from the cell pellet with a small well pipette. (2) Use 200 pL of lysis buffer (20 mM Tris pH 8.0, 125 pg/rnL proteinase K, 5 mM mM NaCl, 10 mM EDTA pH 8.0, 0.5% SDS) resuspended pellets; (3) cell line was dissolved at 5 ° C for 1 hour; (4 dissolved mixture was pipetted to a phase-locked gel (PLG - Eppendorf) 2mL·*# ; (5) Add an equal volume of p:c:l and allow mixing for 5 hours; (6) Centrifuge the official at 12k xg for 5 minutes; (7) Transfer the aqueous phase from the plg tube The gel was removed and an equal volume of gas was added to the aqueous phase and mixed for 30 minutes; (8) The tube was centrifuged at 14k for approximately 5 minutes' (9) the top layer (aqueous phase) Use a pipette to remove the gas, and place it in a new tube at 104 201038734; (10) add 0 J volume of 3M NaOAC and mix (reverse several times); (11) add 2 volumes of 1 inch % EtOH and mixed with the genomic DNA precipitate formed at this stage (inverted several times); (丨2) the tubes are attached to a microcentrifuge tube Rotating at 14k for about 15 minutes at 4t; (13) The remaining gene body dna at the bottom of the liquid system and the tube is gently poured off; (14) Washing the pellet with 〇5mL 70% EtOH; (15) The tube is tied in a microcentrifuge tube at 4. (: at a 14k rotation for about 5 minutes; (16) gently dumping EtOH and drying the genomic DNA pellet; and (17) an appropriate volume of strontium and RNase It is directly added to the genomic DNA pellet. The detached genomic DNA is used to generate a recombinant library consisting of large fragments (approximately 40 kB) (in accordance with the manufacturer's instructions) on the plastid pWEB. -TNCTM (Epicentre). The plastid library was screened by standard community hybridization procedures using 32P radiolabeled probes (Sambrook J. and Russell D. 2001. Molecular cloning: A laboratory manual, 3rd) Edition. Cold Spring Harbor

Laboratory Press, Cold Spring Harbor, New York)。該等探針 含有如實施例1說明的來自其他有機體之公開的PUFA合成 酶序列有同源性的DNA。此等探針係藉由從以上說明的 pJET1.2/純的而由各別的殖株之經選殖的片段之DNA限制 性消化來產生以及藉由標準的方法予以標記。於所有的情 況中，個別的探針之強的雜交至某些黏接質體表示含有與 PUFA合成酶基因同源性的DNA之殖株。 黏接質體殖株PDS115係展現出探針之強的雜交至該 105 201038734 KS區域以及被挑選用於裂瘦壺菌種ATCC PTA-9695 基因的DNA定序。黏接質體殖株pDSll5，其含有裂殖壺菌 種ATCC PTA-%95 與朽沿基因，係依據布達佩斯條 約於2009年1月27日被寄存於美國類型培養物收集中心， 專利寄存 ’ 10801 University Boulevard, Manassas, VA 20110-2209，以及授予ATCC寄存編號PTA-9737。實施例1 中決定的該KS區域之DNA序列的定序引子係使用標準的 方法來設計。為了決定裂殖壺菌種ATCC PTA-9695 /7¾ /的 D N A序列，後繼回合的D N A定序（涉及藉由標準的方法設計 之隨後的定序引子）係予以進行俾以"走動"該黏接質體殖 株。 於先前發表的破囊壺菌PUFA合成酶系統中，PUFA合 成酶基因PFJ/與PFJ2已經被群集在一起且被配置為要分 歧轉錄的。此也是來自裂殖壺菌種ATCC PTA-9695的PE47 與PE42之情況。經由從黏接質體殖株pDS115"走動” DNA 序列’發現户柯2的概念起點是在户柯/的起點之493個核普 酸以及分歧轉錄的。裂殖壺菌種ATCC PTA-9695 PE47與 PUFA合成酶基因的各核苷酸鹼基對係由具有高品質 的至少二個獨立的DNA定序反應來涵括，其具有40之至少 最小的聚集的弗萊德分數(Phred score) (99.99%的信賴位 準）。 黏接質體殖株pBS4係展現出探針之強的雜交至該dh 區域以及被挑選用於裂殖壺菌種ATCC PTA-9695户柯3基 因的DNA定序。含有裂殖壺菌種ATCC PTA-9695 基因 106 201038734 之黏接質體殖株PBS4係依據布達佩斯條約於汕卯年丨月” 曰被寄存於美國類型培養物收集中心，專利寄存，ι〇8〇ιLaboratory Press, Cold Spring Harbor, New York). These probes contain DNA homologous to the disclosed PUFA synthase sequences from other organisms as described in Example 1. These probes were generated by DNA restriction digestion from the pJET1.2/pure described above and from the cloned fragments of the respective colonies and were labeled by standard methods. In all cases, strong hybridization of individual probes to certain adherent plastids indicates a strain containing DNA homologous to the PUFA synthase gene. The adherent plastid PDS115 line exhibited strong hybridization of the probe to the 105 201038734 KS region and DNA sequencing selected for the ATCC PTA-9695 gene. The plastid colony pDSll5, which contains the Schizochytrium strain ATCC PTA-%95 and the decay edge gene, was deposited with the American Type Culture Collection Center on January 27, 2009 under the Budapest Treaty, patent registration '10801 University Boulevard, Manassas, VA 20110-2209, and ATCC registration number PTA-9737. The sequencing primers of the DNA sequence of the KS region determined in Example 1 were designed using standard methods. In order to determine the DNA sequence of the Schizochytrium strain ATCC PTA-9695 /73⁄4 /, the subsequent round of DNA sequencing (involving subsequent sequencing primers designed by standard methods) was carried out by "walking" Adhesive plastids. In the previously published Thraustochytrium PUFA synthase system, the PUFA synthase genes PFJ/ and PFJ2 have been clustered and configured to be transcribed. This is also the case of PE47 and PE42 from the Schizochytrium strain ATCC PTA-9695. The concept starting point of Chuke 2 was found to be the 493 nucleotides at the beginning of Hu Ke/ and the divergent transcription by the pDS115" walking DNA sequence from the plastid colony. Schizontium ATCC PTA-9695 PE47 Each nucleotide base pair with the PUFA synthase gene is encompassed by at least two independent DNA sequencing reactions of high quality having a minimum of 40 aggregated Phred scores ( 99.99% confidence level) The plastid progeny pBS4 line exhibits strong hybridization of the probe to the dh region and DNA sequencing selected for the Schizochytrium sp. ATCC PTA-9695 The viscous plastid PBS4 containing the Schizochytrium sp. ATCC PTA-9695 gene 106 201038734 is deposited under the Budapest Treaty in the United States Type Culture Collection, patent deposit, ι〇8 〇ι

University Boulevard, Manassas，VA 20110-2209，以及授予 ATCC寄存編號PTA_9736。實施例i中決定的該DH區域之 DNA序列的定序引子係使用標準的方法來設計。為了決定 的裂殖壺菌種ATCC PTA-9695户烈3基因之DNA序列，後繼 回合的DNA定序（涉及藉由標準的方法設計之隨後的定序 引子）係被進行俾以”走動”該黏接質體殖株。裂殖壺菌種 ATCC PTA-9695 基因的各核普酸驗基對係由高品質 的至少二個獨立的DNA定序反應來涵括，其具有40之至少 最小的聚集的弗萊德分數(99.99%的信賴位準）。 表1顯示裂殖壺菌種ATCC ΡΤΑ-9695 (序列辨識 編號：1)、户抑2 (序列辨識編號：3)，以及PiMJ (序列辨識 編號：5)多核苷酸序列當與先前發表的序列比較時之同一 性。同一性係藉由DNA排列的標準，VectorNTI程式的 "AlignX"程式中之計分矩陣"swgapdnamt"予以決定。 表1 :對PE4/、，以及尸/¾]多核苷酸序列之百分比同 一性 發表的， 和/7¾ J序列的來源的來源 發表的 PFAl (orfA) 與PE47(序列 辨識編號：1) 的％同一性 發表的 PFA2 {orfB) 與PF42(序列 辨識編號：3) 的°/。同一性 發表的 PFA3 (orfQ 與PFA?(序列 辨識編號：5) 的°/«同一性 裂殖壺菌種ATCC 20888 70 66 75 金黃色破囊壺菌ATCC 34304 65 62 未發表的 破囊壺菌種23B ATCC 20892 56 55 67 107 201038734 表2顯示裂殖壺菌種ATCC PTA-9695 Pfalp (序列辨識 編號：2)、Pfa2p (序列辨識編號：4) ’以及Pfa3p (序列辨識 編號：6)胺基酸序列當與先前發表的PUFA合成酶胺基酸序 列比較時之同一性。同一性係藉由使用蛋白質排列的標 準，VectorNTI程式的"AlignX"程式中之計分矩陣" blosum62mt2’’ 予以決定。 表2 :對Pfalp、Pfa2p以及Pfa3p胺基酸序列之百分比同一性 發表的Pfalp、Pfa2p，以及 Pfa3p岸列的來源 發表的 Pfalp(OrfA) 與Pfalp(序列 辨識編號：2) 的％同一性 發表的 Pfa2p(OrfB) 與Pfa2p(序列 辨識編號：4) 的％同一性 發表的 Pfa3p(OrfC) 與Pfa3p(序列 辨識編號：6) 的％同一性 裂殖壺菌種ATCC 20888 60 53 70 金黄色破囊壺菌ATCC 34304 60 54 未發表的 破囊壺菌種23BATCC 20892 52 52 70 實施例3 執行領域分析以各別地注解裂殖壺菌種ATCC PTA-9695 、PE42，以及的PUFA合成酶領域與活 性位置之序列座標。領域係根據與已知的PUFA合成酶、脂 肪酸合成酶，以及聚酮合成酶領域之同源性來鑑定。 表3顯示與裂殖壺菌種ATCC ΡΤΑ-9695 ΡΜ7關聯的領 域與活性位置。 108 201038734 領域 DNA位置 AA位置 位址 DNA位置 AA位置 KS 序列辨識編號：1的 7-1401 序列辨識編號：2的 3467 活性-DXAC* 序列辨識編號：1 的607~609 序列辨識編號：2 的 C203 (序列辨識編號：7) (序列辨識編號：8) (序列辨識編號:43) End - GFGG 序列辨識編號：1的 1363-1374 序列辨識編號：2 的455458 (序列辨識編號：44) (序列辨識編號:45) MAT 1798-2700 序列辨識編號：2的 600-900 活性GHS*LG 序列辨識編號：1 的2095-2097 序列辨識編號：2 的 S699 (序列辨識編號：9) (序列辨識編號：10) (序列辨識編號:46) ACP 序列辨識編號：1的 3298-5400 序列辨識編號：2的 1100-1800 ACPI領域 序列辨識編號：1的 3325-3600 序列辨識編號：2的 1109-1200 (序列辨識編號：11) (序列辨識編號:12) (序列辨識編號：13) (序列辨識编號：14) ACP丨活性 LGIDS* 序列辨識編號：1 的345Φ3456 序列辨識編號：2 的S1152 (序列辨識編號:47) ACP2領域 序列辨識編號：1的 3667-3942 序列辨識編號：2的 1223-1314 (序列辨識编號：15) (序列辨識編號：16) ACP2活性 LGIDS* 序列辨識編號：1 的3796-3798 序列辨識編號：2 的S1266 (序列辨識編號:47) ACP3領域 序列辨識編號：1的 40154290 序列辨識編號：2的 1339-1430 (序列辨識编號：17) (序列辨識編號：18) ACP3活性 LGIDS* (序列辨識編號:47) 序列辨識編號：1 的 41444146 序列辨識編號：2 的S1382 ACP4領域 序列辨識編號：1的 4363-4638 序列辨識編號：2的 1455-1546 (序列辨識編號：19) (序列辨識編號:2〇) ACP4活性 LGIDS* 序列辨識編號：1 的4492^4494 序列辨識編號：2 的S1498 (序列辨識編號:47) ACP5領域 序列辨識編號：1的 47114986 序列辨識編號：2的 1571-1662 (序列辨識編號:21) (序列辨識編號:22) ACP5活性 LGIDS* 序列辨識編號：1 ^4840-4842 序列辨識編號：2 的S1614 (序列辨識編號:47) ❹ 〇 109 201038734 領域 DNA位置 AA位置 位址 DNA位置 AA位置 ACP6領域 序列辨識編號：1的 5053-5328 (序列辨識編號:23) 序列辨識编號：2的 1685-1776 (序列辨識編號:24) ACP6活性 LGIDS* (序列辨識編號:4乃 序列辨識編號：1 的5182-5184 序列辨識編號：2 的S1728 序列辨識編號：1的 5623-7800 序列辨識編號: 1875-2600 2的 1‘核心區域” 序列辨識編號：1的 5998-6900 序列辨識編號：2的 2000-2300University Boulevard, Manassas, VA 20110-2209, and ATCC registration number PTA_9736. The sequencing primers for the DNA sequence of the DH region determined in Example i were designed using standard methods. In order to determine the DNA sequence of the Schizochytrium sp. ATCC PTA-9695 Hurley 3 gene, subsequent DNA sequencing (involving subsequent sequencing primers designed by standard methods) was performed to "walk around" Adhesive plastids. Each of the nucleocapsid assays of the Schizochytrium sp. ATCC PTA-9695 gene is encompassed by a high quality of at least two independent DNA sequencing reactions with at least a minimum aggregated Fryd score of 40 ( 99.99% confidence level). Table 1 shows that Schizochytrium sp. ATCC ΡΤΑ-9695 (SEQ ID NO: 1), Suppressor 2 (SEQ ID NO: 3), and PiMJ (SEQ ID NO: 5) polynucleotide sequences when compared with previously published sequences The identity of the comparison. Identity is determined by the standard of DNA arrangement, the scoring matrix "swgapdnamt" in the "AlignX" program of the VectorNTI program. Table 1: Percentage identity of the PE4/, and corpse/3⁄4] polynucleotide sequences, and the source of the source of the /73⁄4 J sequence published as % of PFAl (orfA) and PE47 (SEQ ID NO: 1) PFA2 {orfB) published by identity and °/ of PF42 (sequence identification number: 3). Identity published by PFA3 (orfQ and PFA? (SEQ ID NO: 5) °/«Identity Schizochytrium strain ATCC 20888 70 66 75 Thraustochytrium aureum ATCC 34304 65 62 Unpublished Thraustochytrium 23B ATCC 20892 56 55 67 107 201038734 Table 2 shows Schizochytrium species ATCC PTA-9695 Pfalp (SEQ ID NO: 2), Pfa2p (SEQ ID NO: 4) 'and Pfa3p (SEQ ID NO: 6) Amino group The identity of the acid sequence when compared to the previously published PUFA synthase amino acid sequence. The identity is determined by using the standard of protein arrangement, the VectorNTI program's "AlignX" program scoring matrix " blosum62mt2'' Decision 2 Table 2: % identity of Pfalp, Pfa2p, and Pfa3p shoreline published by Pfalp, Pfa2p, and Pfa3p amino acid sequence published by Pfalp (OrfA) and Pfalp (SEQ ID NO: 2) Pone expression of Pfa2p (OrfB) and Pfa2p (SEQ ID NO: 4) % of the identity of Pfa3p (OrfC) and Pfa3p (SEQ ID NO: 6) % identity Schizochytrium species ATCC 20888 60 53 70 golden yellow broken Phytophthora ATCC 34304 60 54 Unpublished Thraustochytrium sp. 23BATCC 20892 52 52 70 Example 3 Performing field analysis to individually annotate Schizochytrium species ATCC PTA-9695, PE42, and PUFA synthetase fields and activities The sequence coordinates of the positions are identified based on homology with known PUFA synthase, fatty acid synthase, and polyketide synthase fields. Table 3 shows the fields associated with Schizochytrium sp. ATCC ΡΤΑ-9695 ΡΜ7. And active position. 108 201038734 Domain DNA position AA position address DNA position AA position KS Sequence identification number: 1 of 7-1401 Sequence identification number: 2 of 3467 Activity - DXAC* Sequence identification number: 1 of 607~609 Sequence identification number C2 of :2 (sequence identification number: 7) (sequence identification number: 8) (sequence identification number: 43) End - GFGG sequence identification number: 1 1363-1374 Sequence identification number: 2 455458 (sequence identification number: 44 (Sequence ID: 45) MAT 1798-2700 Sequence ID: 2 600-900 Active GHS*LG Sequence ID: 1 2095-2097 Sequence ID: 2 S699 (Sequence Identification No.: 9) (Sequence identification number: 10) (Sequence identification number: 46) ACP Sequence identification number: 1 3298-5400 Sequence identification number: 2 1100-1800 ACPI domain sequence identification number: 1 of 3325-3600 Sequence identification No.: 1109-1200 (sequence identification number: 11) (sequence identification number: 12) (sequence identification number: 13) (sequence identification number: 14) ACP 丨 active LGIDS* sequence identification number: 1 345Φ3456 sequence identification No.: S1152 (sequence identification number: 47) ACP2 domain sequence identification number: 1 3667-3942 Sequence identification number: 2 1223-1314 (sequence identification number: 15) (sequence identification number: 16) ACP2 active LGIDS * Sequence identification number: 1 3796-3798 Sequence identification number: 2 S1266 (sequence identification number: 47) ACP3 domain sequence identification number: 1 40154290 Sequence identification number: 2 of 1339-1430 (sequence identification number: 17) (Sequence identification number: 18) ACP3 activity LGIDS* (sequence identification number: 47) Sequence identification number: 1 41444146 Sequence identification number: 2 S1382 ACP4 domain sequence identification No.: 4363-4638 Sequence identification number: 2 1455-1546 (sequence identification number: 19) (sequence identification number: 2〇) ACP4 activity LGIDS* Sequence identification number: 1 of 4492^4494 Serial identification number: 2 S1498 (sequence identification number: 47) ACP5 domain sequence identification number: 1 47114986 Sequence identification number: 2 1571-1662 (sequence identification number: 21) (sequence identification number: 22) ACP5 activity LGIDS* Sequence identification number: 1 ^ 4840-4842 Sequence identification number: 2 S1614 (sequence identification number: 47) ❹ 〇109 201038734 Field DNA position AA position address DNA position AA position ACP6 field sequence identification number: 1 5053-5328 (sequence identification number: 23) Sequence identification number: 1685-1776 of 2 (sequence identification number: 24) ACP6 active LGIDS* (sequence identification number: 4 is sequence identification number: 1 of 5182-5184 sequence identification number: 2 of S1728 sequence identification number: 1 5623-7800 Serial Identification Number: 1875-2600 2's 1' Core Area' Sequence Identification Number: 1 of 5998-6900 Serial Identification Number: 2 of 2000-2300

KR (序列辨識編號:25) (序列辨識編號:26) (序列辨識編號:48) (序列辨識編號:49) DH 序列辨識編號：1的 7027-7065 序列辨識編號：2的 2343-2355 LxxHxxxGxxxxP 序列辨識編號：ΰΓ 7027-7065 序列辨識編號：2的 2343-2355 要素 (序列辨識編號:27) (序列辨識編號:28) (序列辨識编號:50) (序列辨識编號:27) (序列辨識編號:28) 裂殖壺菌種ATCC PTA-9695 Pfal内的第一個領域為 KS領域。含有編碼裂殖壺菌種ATCC PTA-9695 Pfal KS領 域的序列之核苷酸序列於本文中表示為序列辨識編號：7， 其係對應於序列辨識編號：1的位置7-1401。包含裂殖壺菌 種ATCC PTA-9695 Pfal KS領域的胺基酸序列於本文中表 示為序列辨識編號：8，其係對應於序列辨識編號：2的位 置3-467。KS領域含有一活性位置要素：DXAC* (序列辨識 編號：43)，具有對應於序列辨識編號：2的C203之一 *醯基 結合位址。而且，一特性要素係存在於KS領域的終端： GFGG (序列辨識編號：44)，其係對應於序列辨識編號：2 的位置455-458以及序列辨識編號：8的位置453-456。 裂殖壺菌種ATCC PTA-9695 Pfal中的第二個領域為 MAT領域。含有編碼該裂殖壺菌種ATCC PTA-9695 Pfal M AT領域的序列之核苷酸序列於本文中係表示為序列辨識 編號：9，其係對應於序列辨識編號：1的位置1798-2700。 110 201038734 包含裂殖壺菌種ATCC PTA-9695 Pfal MAT領域的胺基酸 序列於本文中係表示為序列辨識編號：10，其係對應於序 列辨識編號：2的位置600-900。MAT領域含有一活性位置 要素：GHS*XG (序列辨識編號：46)，具有對應於序列辨 識編號：2的S699之一 *醯基結合位址。 裂殖壺菌種ATCC PTA-9695 Pfal的第三至第八個領域 為六個串聯ACP領域，於本文中亦稱為ACPI、ACP2、 _ ACP3、ACP4、ACP5，以及 ACP6 〇 含有第一 ACP領域，ACP1 〇 的核苷酸序列於本文中係表示為序列辨識編號：13，以及 包含序列辨識編號：1的大約位置3325延伸到大約位置3600 的核苷酸序列。含有ACPI的胺基酸序列，本文中表示為序 ' 列辨識編號：14，係包含序列辨識編號：2的大約位置1109 ’ 延伸到大約位置1200的胺基酸序列。含有ACP2的核苷酸序 列，本文中表示為序列辨識編號：15，係包含序列辨識編 號：1的大約位置3667延伸到大約位置3942的核苷酸序列。 q 含有ACP2的胺基酸序列，本文中表示為序列辨識編號： 16，係包含序列辨識編號：2的大約位置1223延伸到大約位 置1314的胺基酸序列。含有ACP3的核苷酸序列，本文中表 示為序列辨識編號：17，係包含序列辨識編號：1的大約位 置4015延伸到大約位置4290的核苷酸序列。含有ACP3的胺 基酸序列，本文中表示為序列辨識編號：18，係包含序列 辨識編號：2的大約位置1339延伸到大約位置1430的胺基酸 序列。含有ACP4的核苷酸序列，本文中表示為序列辨識編 號：19，係包含序列辨識編號：1的大約位置4363延伸到大 111 201038734 約位置4638的核苷酸序列。含有ACP4的胺基酸序列，本文 中表示為序列辨識編號：20，係包含序列辨識編號：2的大 約位置1455延伸到大約位置1546的胺基酸序列。含有ACP5 的核苷酸序列，本文中表示為序列辨識編號：21，係包含 序列辨識編號：1的大約位置4711延伸到大約位置4986的核 苷酸序列。含有ACP5的胺基酸序列，本文中表示為序列辨 識編號：22，係包含序列辨識編號：2的大約位置1571延伸 到大約位置1662的胺基酸序列。含有ACP6的核_酸序列， 本文中表示為序列辨識編號：23，係包含序列辨識編號：1 的大約位置5053延伸到大約位置5328的核苷酸序列。含有 ACP6的胺基酸序列，本文中表示為序列辨識編號：24，係 包含序列辨識編號：2的大約位置1685延伸到大約位置1776 的胺基酸序列。全部的六個ACP領域一起延伸裂殖壺菌種 ATCC PTA-9695 Pfal的一區域，從序列辨識編號：1的大約 位置3298至大約位置5400，對應於序列辨識編號：2的大約 1100至大約1800的胺基酸位置。含有全部的6個領域之整個 A C P區域的核苷酸序列於本文中係表示為序列辨識編號： 11 ;而含有全部的6個領域的整個ACP區域之胺基酸序列於 本文中係表示為序列辨識編號：12。序列辨識編號：11内 之6個ACP領域之重複的間隔發現為大概每342個核苷酸(經 測量之鄰近活性位置絲胺酸之間的實際胺基酸數目範圍為 114至116個胺基酸）。6個ACP領域之每一者皆含有一泛硫醇 結合要素LGIDS*(序列辨識編號：47)，其中S*為該泛硫醇 結合位置絲胺酸(S)。該泛硫醇結合位置絲胺酸(S)係位於接 112 201038734 近每一ACP領域序列中心處。6個ACPD領域之每一者的活 性位置絲胺酸殘基的位置（亦即，該泛硫醇結合位置），關於 序列辨識編號：2的胺基酸序列而言，為：ACPI = S1152 ’ ACP2 = S1266，ACP3 = S1382，ACP4 = S1498，ACP5 = S1614，以及ACP6 = S1728。 裂殖壺菌種ATCC PTA-9695 Pfal中的第9個領域為KR 領域。含有編碼裂殖壺菌種ATCC PTA-9695 Pfal KR領域的 序列之核苷酸序列於本文中係表示為序列辨識編號：25， 其係對應於序列辨識編號：1的位置5623-7800。包含裂殖 壺菌種ATCC PTA-9695 Pfal KR領域之胺基酸序列於本文 中係表示為序列辨識編號：26,其係對應於序列辨識編號： 2的位置1875-2600。在KR領域中為一核心區域(係包含於序 列辨識編號：48的核苷酸序列，以及序列辨識編號：49的 胺基酸序列），與短鏈醛類-脫氫酶具有同源性(KR為此家族 之—成員）。此核心區域係自序列辨識編號：1的大約位置 5998延伸至大約6900，其係對應於序列辨識編號：2的胺基 酸位置2000-2300。 裂殖壺菌種ATCC PTA-9695 Pfal中的第10個領域為 DH領域。含有編碼裂殖壺菌種ATcc PTA-9695 Pfal DH領 域的序列之核苷酸序列於本文中係表示為序列辨識編號： 27 ’其係對應於序列辨識編號：1的位置7027-7065。包含 裂殖壺菌種ATCC PTA-9695 Pfal DH領域的胺基酸序列於 本文中係表示為序列辨識編號：28，其係對應於序列辨識 編號：2的位置2343-2355。DH領域含有守恆性活性位置要 113 201038734 素（參見，Donadio, S. and Katz.，L·, Gene 111(1): 51-60 (1992)): LxxHxxxGxxxxP(序列辨識編號：50)。 表4顯示與裂殖壺菌種ATCC PTA-9695 PFA2關聯的領 域與活性位置。 表4:裂殖壺菌種ATCCPTA-9695 PFA2領域分析 領域 DNA位置 AA位置 位址 DMA位置 AA位置 KS 序列辨識編號：3的 10-1350 序列辨識編號：4的 4450 DXAC* 序列辨識編號：3 的 5Ή-573 序列辨識編號：4 的 C191 (序列辨識編號:29) (序列辨識編號:30) (序列辨識編號:43) End-GFGG 序列辨識編號：3的 1312-1323 序列辨識編號：4 的 438·441 (序列辨識編號：44) (序列辨識编號：51) CLF 序列辨識編號：3的 1408-2700 序列辨識編號：4的 470-900 (序列辨識編號:31) (序列辨識編號:32) AT 序列辨識編號：3的 29984200 序列辨識編號：4的 1000-1400 GxS*xG 序列辨識編號：3 的3421-3423 序列辨識編號：4 的 S1141 (序列辨識编號:33) (序列辨識編號:34) (序列辨識編號:52) ER 序列辨識編號：3的 4498-5700 序列辨識編號：4的 1500-1900 (序列辨識編號:3¾ (序列辨識編號:36)KR (sequence identification number: 25) (sequence identification number: 26) (sequence identification number: 48) (sequence identification number: 49) DH sequence identification number: 1 7027-7065 sequence identification number: 2 of 2343-2355 LxxHxxxGxxxxP sequence Identification number: ΰΓ 7027-7065 Sequence identification number: 2 2343-2355 Element (sequence identification number: 27) (sequence identification number: 28) (sequence identification number: 50) (sequence identification number: 27) (sequence identification No.: 28) Schizophrenia strain ATCC PTA-9695 The first field in Pfal is the KS field. The nucleotide sequence containing the sequence encoding the Schizochytrium sp. ATCC PTA-9695 Pfal KS domain is represented herein as sequence identification number: 7, which corresponds to position 7-1401 of sequence identification number: 1. The amino acid sequence comprising the Schizochytrium sp. ATCC PTA-9695 Pfal KS domain is represented herein as Sequence Identification Number: 8, which corresponds to position 3-367 of Sequence Identification Number: 2. The KS field contains an active positional element: DXAC* (sequence identification number: 43) with one of C203 corresponding to sequence identification number: 2 * thiol binding address. Moreover, a characteristic element exists in the terminal of the KS field: GFGG (Sequence Identification Number: 44), which corresponds to the position 455-458 of the sequence identification number: 2 and the position 453-456 of the sequence identification number: 8. The second area of the Schizochytrium strain ATCC PTA-9695 Pfal is the MAT field. The nucleotide sequence containing the sequence encoding the Schizochytrium ATCC PTA-9695 Pfal M AT domain is represented herein as Sequence Identification Number: 9, which corresponds to position 1798-2700 of Sequence Identification Number: 1. 110 201038734 The amino acid sequence comprising the Schizochytrium strain ATCC PTA-9695 Pfal MAT is represented herein as Sequence Identification Number: 10, which corresponds to position 600-900 of Sequence Identification Number: 2. The MAT field contains an active position element: GHS*XG (SEQ ID NO: 46) with one of the S699 corresponding to the sequence identification number: 2 * 醯-based binding address. The third to eighth areas of the Schizochytrium strain ATCC PTA-9695 Pfal are six tandem ACP domains, also referred to herein as ACPI, ACP2, _ ACP3, ACP4, ACP5, and ACP6 〇 containing the first ACP domain. The nucleotide sequence of ACP1 〇 is represented herein as sequence identification number: 13, and a nucleotide sequence comprising sequence identification number: 1 extending from position 3325 to approximately position 3600. The amino acid sequence containing ACPI, designated herein as the sequence 'column identification number: 14, is an amino acid sequence comprising an approximate position 1109' of sequence identification number: 2 extending to approximately position 1200. The nucleotide sequence containing ACP2, designated herein as Sequence Identification Number: 15, is a nucleotide sequence comprising the sequence recognition number: 1 extending from position 3667 to approximately position 3942. q Amino acid sequence containing ACP2, designated herein as Sequence Identification Number: 16, comprises an amino acid sequence of sequence identification number: 2 extending from about position 1223 to about position 1314. The nucleotide sequence containing ACP3, represented herein as sequence identification number: 17, is a nucleotide sequence comprising sequence identification number: 1 extending from position 4015 to position 4290. The amino acid sequence containing ACP3, designated herein as sequence identification number: 18, comprises an amino acid sequence of sequence identification number: 2 extending from position 1339 to position 1430. A nucleotide sequence comprising ACP4, designated herein as a sequence identifier number: 19, comprises a nucleotide sequence of sequence identification number: 1 extending from position 4363 to a large 111 201038734 about position 4638. The amino acid sequence containing ACP4, designated herein as Sequence Identification Number: 20, comprises an amino acid sequence of sequence identification number: 2 extending from about 1455 to about position 1546. The nucleotide sequence containing ACP5, designated herein as Sequence ID: 21, contains a nucleotide sequence of sequence identification number: 1 extending from position 4711 to position 4986. The amino acid sequence containing ACP5, designated herein as Sequence Identification Number: 22, comprises an amino acid sequence extending from about position 1571 of sequence identification number: 2 to about position 1662. The nuclear-acid sequence containing ACP6, represented herein as sequence identification number: 23, comprises a nucleotide sequence of sequence identification number: 1 extending from position 5053 to position 5328. The amino acid sequence containing ACP6, designated herein as Sequence Identification Number: 24, comprises an amino acid sequence of sequence identification number: 2 extending from about position 1685 to about position 1776. All six ACP domains together extend a region of the Schizochytrium strain ATCC PTA-9695 Pfal, from approximately position 3298 of sequence identification number: 1 to approximately position 5400, corresponding to approximately 1100 to approximately 1800 of sequence identification number: 2 Amino acid position. The nucleotide sequence containing the entire ACP region of all six domains is represented herein as the sequence ID: 11; and the amino acid sequence containing the entire ACP region of all six domains is represented herein as a sequence. Identification number: 12. Sequence identification number: Repeated intervals of 6 ACP domains within 11 were found to be approximately every 342 nucleotides (the actual number of amino acids between the measured active sites of serine acid ranged from 114 to 116 amine groups) acid). Each of the six ACP domains contains a panthenol binding element LGIDS* (SEQ ID NO: 47), where S* is the panthenol binding site serine (S). The panthenol binding site serine acid (S) is located at the center of the sequence of each ACP domain near 112 201038734. The position of the active position of the serine residue in each of the six ACPD domains (i.e., the position of the panthenol), with respect to the amino acid sequence of sequence number: 2, is: ACPI = S1152 ' ACP2 = S1266, ACP3 = S1382, ACP4 = S1498, ACP5 = S1614, and ACP6 = S1728. The ninth field of the Schizochytrium strain ATCC PTA-9695 Pfal is the KR field. The nucleotide sequence containing the sequence encoding the Schizochytrium sp. ATCC PTA-9695 Pfal KR domain is represented herein as Sequence Identification Number: 25, which corresponds to position 5623-7800 of Sequence Identification Number: 1. The amino acid sequence comprising the Schizochytrium species ATCC PTA-9695 Pfal KR is represented herein as Sequence Identification Number: 26, which corresponds to position 1875-2600 of Sequence Identification Number: 2. In the KR domain, a core region (containing the nucleotide sequence of sequence identification number: 48, and the amino acid sequence of sequence identification number: 49) has homology to the short-chain aldehyde-dehydrogenase ( KR is a member of this family). This core region extends from approximately position 5998 of sequence identification number: 1 to approximately 6900, which corresponds to the amino acid position 2000-2300 of sequence identification number:2. The 10th field of the Schizochytrium strain ATCC PTA-9695 Pfal is the DH field. The nucleotide sequence containing the sequence encoding the Schizochytrium sp. ATcc PTA-9695 Pfal DH domain is represented herein as the sequence ID: 27 ' corresponds to position 7027-7065 of sequence identification number: 1. The amino acid sequence comprising the Schizochytrium species ATCC PTA-9695 Pfal DH is represented herein as Sequence Identification Number: 28, which corresponds to position 2343-2355 of Sequence Identification Number: 2. The DH domain contains a conserved active position. 113 201038734 (see, Donadio, S. and Katz., L., Gene 111(1): 51-60 (1992)): LxxHxxxGxxxxP (SEQ ID NO: 50). Table 4 shows the domains and active sites associated with Schizochytrium sp. ATCC PTA-9695 PFA2. Table 4: Schizochytrium strain ATCCPTA-9695 PFA2 domain analysis field DNA position AA position address DMA position AA position KS Sequence identification number: 3 10-1350 Sequence identification number: 4 of 4450 DXAC* Sequence identification number: 3 5Ή-573 Sequence identification number: 4 C191 (sequence identification number: 29) (sequence identification number: 30) (sequence identification number: 43) End-GFGG sequence identification number: 3 1312-1323 Sequence identification number: 4 of 438 · 441 (sequence identification number: 44) (sequence identification number: 51) CLF sequence identification number: 3 of 1408-2700 Sequence identification number: 4 of 470-900 (sequence identification number: 31) (sequence identification number: 32) AT sequence identification number: 3, 29,984,200 Sequence identification number: 4, 1000-1400 GxS*xG Sequence identification number: 3, 3421-3423 Serial identification number: 4, S1141 (sequence identification number: 33) (sequence identification number: 34 (Serial Identification Number: 52) ER Sequence Identification Number: 3 4498-5700 Sequence Identification Number: 4 1500-1900 (Sequence Identification Number: 33⁄4 (Serial Identification Number: 36)

裂殖壺菌種ATCC PTA-9695 Pfa2内的第一個領域為 KS領域。含有編碼裂殖壺菌種ATCC PTA-9695 Pfa2 KS領 域的序列之核苷酸序列於本文中係表示為序列辨識編號： 29，其係對應於序列辨識編號：3的位置10-1350。包含裂 殖壺菌種ATCC PTA-9695 Pfa2 KS領域的胺基酸序列於本 文中係表示為序列辨識編號：30，其係對應於序列辨識蝙 號：4的位置4-450。KS領域含有一活性位置要素：DXA〇 (序列辨識編號：43)，具有對應於序列辨識編號：4的C191 114 201038734 之一*醯基結合位址。而且，一特性要素係存在於尺8領域的 終端：GFGG(序列辨識編號：44)，其係對應於序列辨識編 號：4的位置438-441以及序列辨識編號：30的位置435-438。 裂殖壺菌種ATCC PTA-9695 Pfa2内的第3個領域為 CLF領域。含有編碼裂殖壺菌種ATCC PTA-9695 Pfa2 CLF 領域的序列之核苷酸序列於本文中係表示為序列辨識編 號：31，其係對應於序列辨識編號：3的位置1408-2700。 包含裂殖壺菌種ATCC PTA-9695 Pfa2 CLF領域的胺基酸序 列於本文中係表示為序列辨識編號：32，其係對應於序列 辨識編號：4的位置470-900。 裂殖壺菌種ATCC PTA-9695 Pfa2内的第3個領域為AT 領域。含有編碼裂殖壺菌種ATCC PTA-9695 Pfa2 AT領域的 序列的核苷酸序列於本文中係表示為序列辨識編號：33， 其係對應於序列辨識編號：3的位置2998-4200。包含裂殖 壺菌種ATCC PTA-9695 Pfa2 AT領域的胺基酸序列於本文 中係表示為序列辨識編號：34,其係對應於序列辨識編號： 4的位置1000-1400。AT領域含有一活性位置要素GxS*Xg (序列辨識編號：52)，其為醯基轉移酶(AT)蛋白質的特徵， 具有對應於序列辨識編號：4的位置S1141的一活性位置絲 胺酸殘基。The first field within the Schizochytrium strain ATCC PTA-9695 Pfa2 is the KS field. The nucleotide sequence containing the sequence encoding the Schizochytrium sp. ATCC PTA-9695 Pfa2 KS domain is represented herein as Sequence Identification Number: 29, which corresponds to position 10-1350 of Sequence Identification Number: 3. The amino acid sequence comprising the Schizochytrium sp. ATCC PTA-9695 Pfa2 KS domain is represented herein as Sequence Identification Number: 30, which corresponds to position 4-48 of the sequence identification bat: 4 . The KS field contains an active positional element: DXA〇 (SEQ ID NO: 43) with one of the C191 114 201038734 corresponding to the sequence identification number: 4; Moreover, a characteristic element exists in the terminal of the rule 8 field: GFGG (Sequence Identification Number: 44), which corresponds to the position identification code: 4 position 438-441 and the sequence identification number: 30 position 435-438. The third field within the Schizochytrium strain ATCC PTA-9695 Pfa2 is the CLF field. The nucleotide sequence containing the sequence encoding the Schizochytrium sp. ATCC PTA-9695 Pfa2 CLF domain is represented herein as the sequence ID: 31, which corresponds to position 1408-2700 of sequence identification number: 3. The amino acid sequence comprising the Schizochytrium species ATCC PTA-9695 Pfa2 CLF is designated herein as Sequence Identification Number: 32, which corresponds to position 470-900 of Sequence Identification Number: 4. The third field within the Schizochytrium strain ATCC PTA-9695 Pfa2 is the AT field. The nucleotide sequence containing the sequence encoding the Schizochytrium sp. ATCC PTA-9695 Pfa2 AT domain is represented herein as Sequence ID: 33, which corresponds to position 2998-4200 of Sequence ID:3. The amino acid sequence comprising the Schizochytrium sp. ATCC PTA-9695 Pfa2 AT domain is represented herein as Sequence Identification Number: 34, which corresponds to the position of Sequence Identification Number: 4, 1000-1400. The AT domain contains an active positional element GxS*Xg (SEQ ID NO: 52) which is characteristic of a thiotransferase (AT) protein having an active position of a serine residue corresponding to position S1141 of sequence identification number: 4. base.

裂殖壺菌種ATCC PTA-9695 Pfa2内的第4個領域為ER 領域。含有編碼裂殖壺菌種ATCC PTA-9695 Pfa2 ER領域的 序列之核苷酸序列於本文中係表示為序列辨識編號：35， 其係對應於序列辨識編號：3的位置4498-5700。包含Pfa2 ER 115 201038734 領域的胺基酸序列於本文中係表示為序列辨識編號：36， 其係對應於序列辨識編號：4的位置1500-1900。 表5顯示與裂殖壺菌種ATCC PTA-9695 PFA3關聯的領 域與活性位置。 表5 :裂殖壺菌種ATCCPTA-9695 PFA3領域分析 領域 DNA位置 AA位置 位址 DNA位置 AA位置 DH1 序列辨識編號：5 的 1-1350 序列辨識編號：6 的 1*450 FxxH*F 序列辨識編號：5 的 931-933 序列辨識編號：6 的 H310 (序列辨識編號:37) (序列辨識編號:38) (序列辨識編號：53) DH2 序列辨識編號：5 的 1501-2700 序列辨識編號：6 的 501-900 FxxH*F 序列辨識編號：5 的 2401-2403 序列辨識編號：6 的 H801 (序列辨識編號:3¾ (序列辨識編號:40) (序列辨識編號：53) ER 序列辨識編號：5 的 28484200 序列辨識編號：6 的 950-1400 (序列辨識編號:41) (序列辨識編號:42) 裂殖壺菌種ATCC PTA-9695 Pfa3的第1個與第2個領域 為DH領域，於本文中各別地稱為DH1和DH2。含有編碼該 裂殖壺菌種ATCC PTA-9695 Pfa3 DH1領域的序列之核苷酸 序列於本文中係表示為序列辨識編號：37，其係對應於序 列辨識編號：5的位置1-1350。包含裂殖壺菌種ATCC PTA-9695 Pfa3 DH1領域的胺基酸序列於本文中係表示為 序列辨識編號：38，其係對應於序列辨識編號：6的位置 1-45〇。含有編碼裂殖壺菌種ATCC PTA-9695 Pfa3 DH2領域 的序列之核苷酸序列於本文中係表示為序列辨識編號： 39，其係對應於序列辨識編號：5的位置1501-2700。包含 裂殖壺菌種ATCC PTA-9695 Pfa3 DH2領域的胺基酸序列於 本文中係表示為序列辨識編號：40，其係對應於序列辨識 116 201038734 編號：6的位置501-900。該等DH領域含有一活性位置要素： FxxH*F (序列辨識編號：53)。含有DH1之中的活性位置要 素之核苷酸序列係對應於序列辨識編號：5的位置 931-933，而含有DH2之中的活性位置要素之核苷酸序列係 對應於序列辨識編號：5的位置2401 -2403。於該要素FxxH*F 中的該活性位置Η*係根據來自Leesong等人，Structure 4:253-64 (1996)與 Kimber 等人，J Biol Chem. 279:52593-602 (2004)的資料，DH1中的該活性位置Η*對應 於序列辨識編號：6的Η310以及DH2中的該活性位置Η*對 應於序列辨識編號：6的Η801。 裂殖壺菌種ATCC PTA-9695 Pfa3的第3個領域為ER領 域。含有編碼裂殖壺菌種ATCC PTA-9695 Pfa3 ER領域的序 列之核苷酸序列於本文中係表示為序列辨識編號：41，其 係對應於序列辨識編號：5的位置2848-4200。包含裂殖壺 菌種ATCC PTA-9695 Pfa3 ER領域的胺基酸序列於本文中 係表示為序列辨識編號：42，其係對應於序列辨識編號：6 的位置950-1400。 實施例4 設計KS、ER與DH PUFA合成酶領域的簡併性引子俾以 單離來自以ATCC寄存編號PTA-10212寄存的經單離的微生 物，亦知道為破囊壺菌種ATCCPTA-10212之對應序列。The fourth field within the Schizochytrium strain ATCC PTA-9695 Pfa2 is the ER field. The nucleotide sequence containing the sequence encoding the Schizochytrium sp. ATCC PTA-9695 Pfa2 ER domain is represented herein as Sequence ID: 35, which corresponds to position 4498-5700 of Sequence Identification Number: 3. The amino acid sequence comprising the Pfa2 ER 115 201038734 domain is represented herein as sequence identification number: 36, which corresponds to position 1500-1900 of sequence identification number:4. Table 5 shows the domains and active sites associated with Schizochytrium sp. ATCC PTA-9695 PFA3. Table 5: Schizochytrium strain ATCCPTA-9695 PFA3 field analysis field DNA position AA position address DNA position AA position DH1 Sequence identification number: 5 1-1350 Sequence identification number: 6 of 1*450 FxxH*F Sequence identification number :5 931-933 Sequence ID: 6 H310 (Sequence ID: 37) (Sequence ID: 38) (Sequence ID: 53) DH2 Sequence ID: 5 1501-2700 Serial ID: 6 501-900 FxxH*F Sequence Identification Number: 5 2401-2403 Serial Identification Number: 6 H801 (Sequence Identification Number: 33⁄4 (Sequence Identification Number: 40) (Sequence Identification Number: 53) ER Serial Identification Number: 5 of 28484200 Sequence identification number: 950-1400 of 6 (sequence identification number: 41) (sequence identification number: 42) The first and second fields of Schizochytrium sp. ATCC PTA-9695 Pfa3 are DH fields, each in this paper Also referred to as DH1 and DH2. The nucleotide sequence containing the sequence encoding the Schizochytrium ATCC PTA-9695 Pfa3 DH1 domain is represented herein as Sequence Identification Number: 37, which corresponds to the sequence identification number: Position 5 of 1-1350. Contains The amino acid sequence of the field of ATCC PTA-9695 Pfa3 DH1 is represented herein as sequence identification number: 38, which corresponds to position 1-45 of sequence identification number: 6. Contains the code for Schizochytrium The nucleotide sequence of the sequence of the ATCC PTA-9695 Pfa3 DH2 domain is represented herein as the sequence identification number: 39, which corresponds to the position of the sequence identification number: 5, 1501-2700. Contains the Schizochytrium strain ATCC PTA -9695 Pfa3 The amino acid sequence of the DH2 domain is represented herein as sequence identification number: 40, which corresponds to position identification 116 201038734 number: 6 position 501-900. These DH domains contain an active positional element: FxxH *F (SEQ ID NO: 53). The nucleotide sequence containing the active position element in DH1 corresponds to the position identification number: 5, position 931-933, and the nucleotide containing the active position element in DH2 The sequence corresponds to position 2401 - 2403 of sequence identification number: 5. The active position in the element FxxH*F is based on from Leesong et al., Structure 4: 253-64 (1996) and Kimber et al., J. Biol Chem. 279:52593-602 ( According to 2004), the active position Η* in DH1 corresponds to Η310 of sequence identification number: 6 and the active position D* in DH2 corresponds to Η801 of sequence identification number:6. The third field of Schizochytrium strain ATCC PTA-9695 Pfa3 is the ER domain. The nucleotide sequence containing the sequence encoding the Schizochytrium sp. ATCC PTA-9695 Pfa3 ER domain is represented herein as sequence identification number: 41, which corresponds to position 2848-4200 of sequence identification number: 5. The amino acid sequence comprising the Schizochytrium strain ATCC PTA-9695 Pfa3 ER is referred to herein as Sequence Identification Number: 42, which corresponds to position 950-1400 of Sequence Identification Number: 6. Example 4 Designing the degeneracy primers in the field of KS, ER and DH PUFA synthase to isolate the isolated microorganisms deposited under ATCC accession number PTA-10212, also known as the Thraustochytrium species ATCCPTA-10212 Corresponding sequence.

破囊壺菌種ATCCPTA-10212P7M/的KS區域（亦即，含 有KS領域的區域）的簡併性引子係根據裂殖壺菌種ATCC 2〇888、金黄色破囊壺菌（ATCC 343〇4)，以及破囊壺菌種23B 117 201038734 ATCC 20892之公開的尸E47(先前稱為orfA或ORF 1)序列來 設計： prDS233 (川貝向）： TGATATGGGAGGAATGAATTGTGTNGTNGAYGC (序 列辨識編號：123) prDS235 (反向）： TTCCATAACAAAATGATAATTAGCTCCNCCRAANCC (序列辨識編號：124)。 破囊壺菌種ATCC ΡΤΑ-1〇212尸/¾2的ER區域（亦即’含 有ER領域的區域）的簡併性引子係根據曰本沙雷菌的序 列、裂殖壺菌種ATCC 20888、金黄色破囊壺菌（ATCC 34304)，以及破囊壺菌種23B ATCC 20892之公開的户柯2(先 前稱為orfB或ORF2)序列來設計：The degenerate primer of the KS region of the Thraustochytrium sp. ATCCPTA-10212P7M/ (ie, the region containing the KS domain) is based on the Schizochytrium strain ATCC 2〇888, Thraustochytrium aureum (ATCC 343〇4) ), and the sequence of the corpse E47 (formerly known as orfA or ORF 1) disclosed by the genus Thraustochytrium 23B 117 201038734 ATCC 20892: prDS233 (Chaobei direction): TGATATGGGAGGAATGAATTGTGTNGTNGAYGC (SEQ ID NO: 123) prDS235 (reverse) ): TTCCATAACAAAATGATAATTAGCTCCNCCRAANCC (Sequence Identification Number: 124). The degenerative primers of the Thraustochytrium strain ATCC ΡΤΑ-1〇212 nectar/3⁄42 ER region (ie, the region containing the ER domain) are based on the sequence of S. salivarius, Schizochytrium sp. ATCC 20888, Designed by the sequence of Thraustochytrium sclerophylla (ATCC 34304) and the genus Cocoa 2 (formerly known as orfB or ORF2) of Thraustochytrium sp. 23B ATCC 20892:

prDS183(順向）：GGCGGCCACACCGAYAAYMGNCC (序列辨識編號：125) prDS 184(反向）：CGGGGCCGCACCANAYYTGRTA (序 列辨識編號：126)。 破囊壺菌種ATCCPTA-l〇2l2Pi^3的ER區域(亦即’含 有ER領域的區域)的簡併性引子係根據日本沙雷菌、裂殖壺 菌種ATCC 20888、金黄色破囊壺菌（ATCC 34304)，以及破 囊壺菌種23B ATCC 20892之公開的尸Λ43(先前稱為orfC或 ORF 3)序列來設計： prDS181(順向）：TCCTTCGGNGCNGSNGG (序列辨識 編號：127) 118 201038734 prDS 184(反向）：CGGGGCCGCACCANAYYTGRTA (序 列辨識編號：126)。 如以上說明之JGM190(順向，序列辨識編號：64)與 BLR242 (反向，序列辨識編號·· 65)的簡併性引子係用來擴 增來自破囊壺菌種ATCC PTA-10212的户柯3之DH區域。 染色體DNA模板之PCR條件係如下：0·2 μΜ dNTPs、 0·1 uM各引子、6% DMSO、200 ng 染色體DNA、2.5 U Herculase® II融合聚合酶（Stratagene)，以及 IX Herculase® 缓衝液(Stratagene)配於50pL總體積中。PCR流程包括以下 步驟：（1) 98°C歷時3分鐘；（2) 98°C歷時30秒；（3) 54°C歷 時45秒；（4) 72°C歷時1分鐘；（5)重複步驟2-4計40個循環； (6)72°C歷時5分鐘；以及⑺維持在6°C。 關於全部的引子對，PCR使用來自破囊壺菌種ATCC PTA-10212之染色體模板出產具有預期的大小之有區別的 DNA產物。各別的PCR產物係依據製造者之指示而被選殖 至該載體pJET1.2/純的（Fermentas)之内，以及使用所提供的 標準引子來決定插入序列。 從PCR產物得到的DNA序列係如實施例1說明的與 NCBI GenBank可得的已知序列比較。 於胺基酸的位準，與含有來自破囊壺菌種ATCC PTA-10212的PEiiiKS片段之選殖的DNA所衍生之演繹的 胺基酸序列具有最高位準的同源性之序列為：裂殖壺菌種 ATCC 20888 "多不飽和脂肪酸合成酶次單元A"(同一性= 80%，陽性=90%);深海沙雷菌〜咐/n’ca) KT99 119 201038734 "亞米加-3多不飽和脂肪酸合成酶PfaA”（同一性=51%;陽性 =67%);沙雷菌（57^冰<3辦//« /ozTz/ca) PV-4 ’'石-酮基驢基合 成酶"（同一性=50% ;陽性=67%);木質沙雷菌（幼6而邮心 woo咖·）ATCC 51908 "聚酮類多不飽和脂肪酸合成酶pfaA" (同一性=51% ;陽性=66%)。 於胺基酸位準’與含有來自破囊壺菌種ATCC PTA-10212的/之ER片段之選殖的DNA所衍生之演繹的 胺基酸序列具有最高位準的同源性之序列為：裂殖壺菌種 ATCC 20888"多不飽和脂肪酸合成酶次單元同一性= 70% ;陽性=85%);裂殖壺菌種ATCC 20888，，多不飽和脂肪 酸合成酶次單元C"(同一性=66% ;陽性=83%);泡沫節球 藻（iVodM/aria vwmigewa) CCY9414 "2-硝基丙烧二氧酶" (同一性=57°/〇 ;陽性=74%);莫里特拉菌物種PE36 "多不 飽和脂肪酸合成酶PfaD"(同一性=57% ;陽性=71%)。prDS183 (forward): GGCGGCCACACCGAYAAYMGNCC (SEQ ID NO: 125) prDS 184 (reverse): CGGGGCCGCACCANAYYTGRTA (serial identification number: 126). The degenerate primer of the ER region of the Thraustochytrium strain ATCCPTA-l〇2l2Pi^3 (that is, the region containing the ER domain) is based on the Japanese Serratia, Schizochytrium ATCC 20888, and the golden yellow sac The bacterium (ATCC 34304), and the sequence of the corpse 43 (formerly known as orfC or ORF 3) disclosed by Thraustochytrium sp. 23B ATCC 20892: prDS181 (forward): TCCTTCGGNGCNGSNGG (SEQ ID NO: 127) 118 201038734 prDS 184 (reverse): CGGGGCCGCACCANAYYTGRTA (sequence identification number: 126). The degenerate primers for JGM190 (forward, sequence identification number: 64) and BLR242 (reverse, sequence identification number 65) as described above are used to amplify households from the Thraustochytrium sp. ATCC PTA-10212 The DH area of Ke 3. The PCR conditions for chromosomal DNA templates are as follows: 0·2 μΜ dNTPs, 0·1 uM primers, 6% DMSO, 200 ng chromosomal DNA, 2.5 U Herculase® II fusion polymerase (Stratagene), and IX Herculase® buffer ( Stratagene) is formulated in a total volume of 50 pL. The PCR procedure consists of the following steps: (1) 98 ° C for 3 minutes; (2) 98 ° C for 30 seconds; (3) 54 ° C for 45 seconds; (4) 72 ° C for 1 minute; (5) repeat Steps 2-4 count 40 cycles; (6) 72 ° C for 5 minutes; and (7) maintained at 6 ° C. For all primer pairs, PCR uses a chromosomal template from Thraustochytrium sp. ATCC PTA-10212 to produce a differentiated DNA product of the expected size. The individual PCR products were cloned into the vector pJET1.2/Fermentas according to the manufacturer's instructions, and the inserted primers were used to determine the insertion sequence. The DNA sequence obtained from the PCR product was compared to the known sequence available in NCBI GenBank as described in Example 1. The sequence with the highest level of homology to the deduced amino acid sequence derived from the DNA of the selected clone of the PEiiiKS fragment from the Thraustochytrium sp. ATCC PTA-10212 is: The genus ATCC 20888 " polyunsaturated fatty acid synthase subunit A" (identity = 80%, positive = 90%); deep sea serratia ~ 咐 / n'ca) KT99 119 201038734 " -3 polyunsaturated fatty acid synthase PfaA" (identity = 51%; positive = 67%); Serratia (57 ^ ice < 3 office / / « / ozTz / ca) PV-4 '' stone - ketone驴 合成 synthase " (identity = 50%; positive = 67%); wood Serratia (young 6 and post woo café) ATCC 51908 " polyketone polyunsaturated fatty acid synthase pfaA" Identity = 51%; positive = 66%). Deduced amino acid sequence derived from amino acid-classified DNA containing the ER fragment from the Thraustochytrium sp. ATCC PTA-10212 The sequence with the highest level of homology is: Schizochytrium strain ATCC 20888" polyunsaturated fatty acid synthase subunit identity = 70%; positive = 85%); Schizochytrium strain ATCC 20888, more Unsaturated fat Acid synthase subunit C" (identity = 66%; positive = 83%); IVodM/aria vwmigewa CCY9414 " 2-nitropropane dioxygenase" (identity = 57°) /〇; positive = 74%); Moritrabacter species PE36 " polyunsaturated fatty acid synthase PfaD" (identity = 57%; positive = 71%).

於胺基酸位準’與含有來自破囊壺菌種ATCC Ρ ΤΑ -10 212的户柯3之E R片段之選殖的DN A所衍生之演繹的 胺基酸序列具有最高位準的同源性之序列為：裂殖壺菌種 ATCC 20888 "多不飽和脂肪酸合成酶次單元c”（同一性= 80%;陽性=90%);裂殖壺菌種ATCC2〇888 ”多不飽和脂肪 酸合成酶次單元(同一性=78% ;陽性=89%);莫里特拉 菌物種PE36"多不飽和脂肪酸合成酶PfaD”（同一性=56〇/〇 ; 以性_ 71/ό) ’ 亞馬遜沙雷菌（灿SB2B ”亞米加-3多不飽和脂肪酸合成酶pfaD"(同一性=55%;陽性 =73%) ° 120 201038734 於胺基酸位準，與含有來自破囊壺菌種ATCC PTA-10212的之DH片段之選殖的DNA所衍生之演繹 的胺基酸序列具有最高位準的同源性之序列為：裂殖壺菌 種ATCC 20888 π多不飽和脂肪酸合成酶次單元C”（同一性= 63% ;陽性=76%);沙雷菌（She而《e//a pea/e⑽a) ATCC 700345 "Β-羥基醯基-(醯基-載體-蛋白質）脫水酶 FabA/FabZn(同一性=35% ;陽性=53%);沙雷菌（57zewa«e//a WP3 "多領域/3-酮基醯基合成酶”（同一性= 36% ;陽性=52%);深海沙雷菌KT99 ”亞米加_3多不飽和 脂肪酸合成酶PfaC"(同一性=35% ;陽性=51%)。 實施例5 PUFA合成酶基因係被鑑定來自破囊壺菌種atcC PTA-10212。 由-80°C冷凍管，1 mL的細胞係在室溫下予以解凍且被 添加至250 mL無檔板燒瓶中的50 mL的液體HSFM培養基 (below)。該燒瓶係在23 °C下孵育歷時3天。收集細胞以及用 於標準的細菌人工染色體（BAC)庫建構（LucigenThe deduced amino acid sequence derived from the amino acid sequence at the amino acid level and the DN A containing the ER fragment of the genus Teke 3 from the Thraustochytrium sp. ATCC Ρ ΤΑ -10 212 has the highest level of homology. The sequence of sex is: Schizochytrium strain ATCC 20888 " polyunsaturated fatty acid synthase subunit c" (identity = 80%; positive = 90%); Schizochytrium strain ATCC2 〇 888" polyunsaturated fatty acid Synthetic enzyme subunit (identity = 78%; positive = 89%); Moritrabacter species PE36 " polyunsaturated fatty acid synthase PfaD" (identity = 56 〇 / 〇; sex _ 71 / ό) ' Serratia marcescens (can SB2B) sub-male-3 polyunsaturated fatty acid synthase pfaD" (identity = 55%; positive = 73%) ° 120 201038734 at amino acid level, with containing from Thraustochytrium The sequence of the deduced amino acid sequence derived from the DNA of the DH fragment of ATCC PTA-10212 has the highest level of homology: Schizochytrium sp. ATCC 20888 π polyunsaturated fatty acid synthase Unit C" (identity = 63%; positive = 76%); Serratia (She and "e//a pea/e(10)a) ATCC 700345 " Β-hydroxy fluorenyl-(mercapto-vector -protein) dehydratase FabA/FabZn (identity = 35%; positive = 53%); Serratia (57zewa «e//a WP3 " multi-domain / 3-ketothiol synthase" (identity = 36%; positive = 52%); Serratia marcescens KT99 ”Amigato _3 polyunsaturated fatty acid synthase PfaC" (identity = 35%; positive = 51%). Example 5 PUFA synthase gene system Identification from the Thraustochytrium strain atcC PTA-10212. From a -80 ° C cryotube, 1 mL of cell line was thawed at room temperature and added to 50 mL of liquid HSFM medium in a 250 mL unflanked flask ( The flask was incubated at 23 ° C for 3 days. Collected cells and used for standard bacterial artificial chromosome (BAC) library construction (Lucigen

Corporation, Middleton, WI USA)。 121 表6 : HSFM培養基 成分 濃度 範圍Corporation, Middleton, WI USA). 121 Table 6: HSFM medium Composition Concentration Range

Na2S04 g/L 31.0 NaCl g/L 0.625 KC1 g/L 1.0 MgS04.7H20 g/L 5.0 (NH4)2S04 g/L 0.44 msg*ih2o g/L 6.0 CaCl2 g/L 0.29 T 154(酵母菌萃取物） g/L 6.0 KH2P〇4 g/L 0.8 高壓蒸氣滅菌後(金屬） 檸檬酸 mg/L 3.5 FeS04_7H20 mg/L 10.30 MnCl2.4H20 mg/L 3.10 ZnS04*7H20 mg/L 3.10 CoC12-6H20 mg/L 0.04 Na2Mo04_2H20 mg/L 0.04 CuS04*5H20 mg/L 2.07 NiS046H20 mg/L 2.07 高壓蒸氣滅菌後(維生素） 硫胺素 mg/L 9.75 維生素B12 mg/L 0.16 Cat泛酸 mg/L 2.06 生物素 mg/L 3.21 高壓蒸氣滅菌後(破） 甘油 g/L 30.0 氮進料： 成分 濃度 MSG_1H20 g/L 17 0-50、15-45，或是 25-35 0-25、0.1-10，或是 0.5-5 0-5、0.25-3，或是 0.5-2 0-10、2-8，或是 3-6 0-10、0.25-5，或是 0.05-3 0-10、4-8，或是 5-7 0.1-5、0.15-3，或是 0.2-1 0-20、0.1-10，或是 1-7 0.1-10、0.5-5，或是 0.6-1.8 0.1-5000、10-3000，或 3-2500 0.1-100、1-50，或 5-25 0.1-100、1-50，或 2-25 0.01-100、1-50，或 2-25 0-1、0.001-0.1，或 0.01-0.1 0.001-1、0.005-0.5，或 0_01-0.1 0.1-100 、 0.5-50 ，或 1-25 0.1-100、0.5-50，或 1-25 0.1-100、1-50，或是 5-25 0.01-100、0.05-5，或是 0.1-1 0.1-100、0.1-50，或是 1-10 0.]-]00、0.1-50，或是 1-10 5-150、10-100，或是 20-50 0-150、10-100，或是 15-50 典型的培養條件會包括以下··Na2S04 g/L 31.0 NaCl g/L 0.625 KC1 g/L 1.0 MgS04.7H20 g/L 5.0 (NH4)2S04 g/L 0.44 msg*ih2o g/L 6.0 CaCl2 g/L 0.29 T 154 (yeast extract) g/L 6.0 KH2P〇4 g/L 0.8 After autoclaving (metal) citrate mg/L 3.5 FeS04_7H20 mg/L 10.30 MnCl2.4H20 mg/L 3.10 ZnS04*7H20 mg/L 3.10 CoC12-6H20 mg/L 0.04 Na2Mo04_2H20 mg/L 0.04 CuS04*5H20 mg/L 2.07 NiS046H20 mg/L 2.07 After autoclaving (vitamin) Thiamine mg/L 9.75 Vitamin B12 mg/L 0.16 Catpantoic acid mg/L 2.06 Biotin mg/L 3.21 High pressure After steam sterilization (broken) Glycerol g/L 30.0 Nitrogen feed: Component concentration MSG_1H20 g/L 17 0-50, 15-45, or 25-35 0-25, 0.1-10, or 0.5-5 0- 5, 0.25-3, or 0.5-2 0-10, 2-8, or 3-6 0-10, 0.25-5, or 0.05-3 0-10, 4-8, or 5-7 0.1-5, 0.15-3, or 0.2-1 0-20, 0.1-10, or 1-7 0.1-10, 0.5-5, or 0.6-1.8 0.1-5000, 10-3000, or 3- 2500 0.1-100, 1-50, or 5-25 0.1-100, 1-50, or 2-25 0.01-100, 1-50, or 2-25 0-1, 0.001-0.1, or 0.01-0.1 0.001 - 1, 0.005-0.5, or 0_01-0.1 0.1-100, 0.5-50, or 1-25 0.1-100, 0.5-50, or 1-25 0.1-100, 1-50, or 5-25 0.01-100 , 0.05-5, or 0.1-1 0.1-100, 0.1-50, or 1-10 0.]-]00, 0.1-50, or 1-10 5-150, 10-100, or 20 -50 0-150, 10-100, or 15-50 Typical culture conditions will include the following ··

PH 溫度： 溶氧： 甘油控制的@ : 約6.5—約9.5、約6.5-約8.0，或約6.8—約 7.8 ； 攝氏約15-約30度、攝氏約18-約28度， 或是攝氏約21約23度； 約0.1-約100%飽和、約5-約50%飽和，或 是約10-約30%飽和；及/或 約5-約50 g/L、約10-約40 g/L，或是約15-約35 g/L. 122 201038734 由大片段（大概120 kB的平均數）所組成的重組型bAC 庫係依據製造者之指示而於BAC载體pSMART® (Lucigen Corporation)中操作。BAC庫係藉由使用32P放射性標記的探 針之4示準的群洛雜父程序予以篩選（Sambr〇〇k J. and Russell D. 2001. Molecular cloning: A laboratory manual, 3rd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York)。該等探針含有如實施例4說明的 來自其他有機體之公開的PUFA合成酶序列有同源性的 DNA。此等探針係藉由從以上說明的pjET1.2/鈍的而由各 別的殖株之經選殖的片段之DNA限制性消化來產生以及藉 由標準的方法予以標記。於所有的情況中，個別的探針之 強的雜交至某些BACs表示含有與PUFA合成酶基因同源性 的DNA之殖株。 BAC殖株pLR 130(亦知道為LuMaBAC 2M23)係展現出 探針之強的雜交至KS區域與ER區域二者，其係指示其含有 PFJ/與PE42基因，以及被挑選用於破囊壺菌種ATCC ΡΤΑ-10212ΡΕ4^/與PE42基因的DNA定序。BAC係藉由標準 的程序予以定序(Eurofins MWG Operon, Huntsville，AL)。 BAC殖株pLR130,其含有與基因，係依據布達佩 斯條約於2009年12月1日被寄存於美國類型培養物收集中 心’專利寄存，10801 University Boulevard, Manassas, VA 20110-2209，以及授予ATCC寄存編號PTA-10511。 於先前發表的破囊壺菌PUFA合成酶系統中，PUFA合 成酶基因尸/¾ /與PE4 2已經被群集在一起且被配置為要分 123 201038734 歧轉錄的。此也是來自破囊壺菌種ATCC PTA-10212的/ 與i5柯2之情況。發現/»柯2的概念起點是在的起點之 693個核苷酸以及分歧轉錄的。 BAC殖株pDS127 (亦知道為LuMaBAC 9K17)係展現出 探針之強的雜交至的DH區域與ER區域二者，以及被 挑選用於/基因的DNA定序。BAC殖株pDS127，其含有 尸柯3基因’係依據布達佩斯條約於2009年12月1日被寄存於 美國類型培養物收集中心，專利寄存，10801 University Boulevard，Manassas, VA 20110-2209，以及授予 ATCC寄存 編號PTA-10510。實施例4中決定的DH區域與ER區域之DNA 序列的定序引子係使用標準的方法來設計。為了決定破囊壺 菌種ATCC PTA-10212 基因的DNA序列，後繼回合的 DNA定序（涉及藉由標準的方法設計之隨後的定序引子）係 被進行俾以”走動”該BAC殖株。基因的各核苷酸鹼基對 係由高品質的至少二個獨立的DNA定序反應來涵括，其具 有40之至少最小的聚集的弗萊德分數(99.99%的信賴位準）。 表7顯示破囊壺菌種ATCC ΡΤΑ-1〇212 序列辨識 編號：68)、序列辨識編號：70)，以及/>似3(序列辨 識編號：72)多核苷酸序列當與先前發表的序列及來自裂殖 壺菌種ΡΤΑ-9695的序列比較時之同一性。同一性係藉由 DNA排列的標準，VectorNTI程式的"AlignX"程式中之計分 矩陣"swgapdnamt"予以決定。 124 201038734 表7 :對户柯/、PE42，以及/>柯3多核苷酸序列之百分比同 一性 比較PFA、PF/I2，以及 PFA?序列的來源 比較PFd/ {orfA)^PFAl 的％同一性 比較 (or/B)與 RFA2 的％同一性 比較 (〇r/QMPFA3 的％同一性 裂殖壺菌種ATCC 20888 55 54 59 金黄色破囊壺菌ATCC 34304 55 53 未發表的 破囊壺菌種23B ATCC 20892 55 57 62 裂殖壺菌種PTA-9695 55 52 59 表8顯示破囊壺菌種ATCC PTA-10212 Pfalp(序列辨識 編號：69)、Pfa2p(序列辨識編號：71)，以及Pfa3p(序列辨 識編號：73)胺基酸序列當與先前發表的PUFA合成酶胺基 酸序列及來自裂殖壺菌種PTA-9695的序列比較時之同一 性。同一性係藉由使用蛋白質排列的標準，VectorNTI程式 的”AlignX1'程式中之計分矩陣”blosum62mt2"予以決定。 表8 :與Pfalp、Pfa2p，以及Pfa3p胺基酸序列之百分比同一性 比較Pfalp, Pfa2p，以及 Pfa3p專列的來源 比較Pfalp (OrfA)與 同 一*lk 比較Pfa2p (OrfB)與 Pfa2p^j% 同一 比較Pfa3p (OrfC)與 卩吻的％ 同一ΊΪ 裂殖壺菌種ATCC 20888 62 57 69 金黄色破囊壺菌ATCC 34304 58 54 未發表的 破囊壺菌種23B ATCC 20892 54 54 71 裂殖壺菌種PTA-9695 59 53 73 實施例6 執行領域分析以各別地注解裂殖壺菌種ATCC PTA-10212 PFW、，以及合成酶領域與 125 201038734 活性位置之序列座標。領域係根據與已知的PUFA合成酶、 脂肪酸合成酶’以及聚酮合成酶領域之同源性來鑑定。 表9顯示與裂殖壺菌種ATCC PTA-10212尸E4/關聯的 領域與活性位置。 表9 :破囊壺菌種ATCC PTA-10212户柯/領域分析 領域 DNA位置 ΑΑ位置 位址 DNA位置 ΑΑ位置 KS 序列辨識編號：68 的 13-1362 序列辨識編號：69 的 5-545 活性-DXAC» 序列辨識編號：68 的 601~612 序列辨識編號：69 的 C204 (序列辨識編號:74) (序列辨識編號:75) (序列辨識編號:43) End-GFGG 序列辨識編號：68 的 1351-1362 序列辨識編號：69 的 451454 (序列辨識编號：44) (序列辨識編號:45) MAT 序列辨識編號：68 的丨 783-2703 序列辨識編號：69 的 595-901 活性GHSIG (序列辨識編號:46) 序列辨識編號：68 的 2083-2085 (鼻列辨識編號： 116) 序列辨識編號：69 的 S695 ACP 序列辨識編號：68 的 3208·6510 (序列辨識編號:78) 序列辨識編號：69 的 1070-2170 (序列辨識編號:79) ACPI領域 序列辨識編號：68 的 3280-3534 (序列辨識編號:80) 的 109Φ1178 (序列辨識編號:81) ACPI活性 LGIDS* (序列辨識·編號:47) 序列辨識編號：68 的 3403-3405 序列辨識編號：69 的 S1135 ACP2領域 序列辨識編號：68 的 3607-3861 序列辨識編號：69 的 1203-1287 (序列辨識編號:8¾ (序列辨識編號:83) ACP2活性 LGIDS* 序列辨識編號··胡 的 3730·3732 序列辨識編號：69 的 S1244 (序列辨識編號:47) ACP3領域 序列辨識編號：68 的 39344185 序列辨識編號：69 的 1312-1396 (序列辨識編號：84) (序列辨識編號：85) ACP3活性 LGIDS* 序列辨識編號：68 的 40574059 序列辨識編號：69 的 S1353 (序列辨識編號:47) ACP4領域 序列辨識編號：68 的 42614515 序列辨識編號：69 的 1421-1505 (序列辨識編號：86) (序列辨識编號:87) ACP4活性 LGIDS* 序列辨識編號：68 的 43844386 序列辨識編號：69 的 S1462 (序列辨識編號:47) 126 201038734 Ο ο 領域 DNA位置 ΑΑ位置 位址 DNA位置 AA位置 ACP5領域 序列辨識編號：68 的 4589*4842 序列辨識編號：69 的 1530-1614 (序列辨識編號:88) (序列辨識編號：89) ACP5活性 LGIDS* 序列辨識编號：68 的 47114713 序列辨識編號：69 的 S1571 f戽列辨識編號:47) ACP6領域 序列辨識編號：68 的 4915-5169 序列辨識編號：69 的 1639-1723 (序列辨識編號:90) (序列辨識編號:91) ACP6活性 LGIDS* 序列辨識編號：68 的 5038-5040 序列辨識編號：69 的 S1680 (庠列辨識編號:47) ACP7領域 序列辨識編號：68 的 5242-54% 序列辨識編號：69 的 1748-1832 (序列辨識編號:92) (序列辨識編號:93) ACP7活性 LGIDS* 序列辨識編號：68 的 5365-5367 序列辨識編號：69 的 S1789 (庠列辨識編號:47) ACP8領域 序列辨識編號：68 的 5569-5823 (序列辨識編號:94) 序列辨識編號：69 的 1857-1941 (序列辨識編號:95) ACP8活性 LGIDS* 序列辨識編號：68 的 5692-5694 序列辨識編號：69 的 S1898 (序列辨識編號:47) ACP9領域 序列辨識編號：68 的 58964150 序列辨識編號：69 的 1966-2050 (序列辨識編號:96) (序列辨識編號:97) ACP9活性 LGIDS* 序列辨識編號：68 的 6019^6021 序列辨識編號：69 的 S2007 (序列辨識編號:47) ACP10領域 序列辨識編號：68 的 6199~6453 序列辨識編號：69 的 2067-2151 (序列辨識編號:98) (序列辨識編號:99) ACT10活性 LGIDS* 序列辨識編號：68 的 63224324 序列辨識編號：69 的 S2108 (序列辨識編號:47) "核心區域"PH temperature: dissolved oxygen: glycerol controlled @: from about 6.5 to about 9.5, from about 6.5 to about 8.0, or from about 6.8 to about 7.8; from about 15 to about 30 degrees Celsius, from about 18 to about 28 degrees Celsius, or about Celsius 21 about 23 degrees; from about 0.1 to about 100% saturated, from about 5 to about 50% saturated, or from about 10 to about 30% saturated; and/or from about 5 to about 50 g/L, from about 10 to about 40 g/ L, or about 15 to about 35 g/L. 122 201038734 A recombinant bAC library consisting of large fragments (approximately 120 kB average) is based on the manufacturer's instructions in the BAC vector pSMART® (Lucigen Corporation) In operation. The BAC library was screened by using the 32P radiolabeled probe 4 as shown in the group's sequence of the father's program (Sambr〇〇k J. and Russell D. 2001. Molecular cloning: A laboratory manual, 3rd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York). These probes contain DNA having homology to the disclosed PUFA synthase sequences from other organisms as described in Example 4. These probes were generated by DNA restriction digestion of the fragments selected from the respective strains of pjET1.2/blunt as explained above and labeled by standard methods. In all cases, strong hybridization of individual probes to certain BACs indicates a strain containing DNA homologous to the PUFA synthase gene. The BAC strain pLR 130 (also known as LuMaBAC 2M23) exhibits strong hybridization of the probe to both the KS region and the ER region, which is indicated to contain the PFJ/PE42 gene, and was selected for Thraustochytrium DNA sequencing of ATCC ΡΤΑ-10212ΡΕ4^/ with PE42 gene. BAC is sequenced by standard procedures (Eurofins MWG Operon, Huntsville, AL). The BAC strain pLR130, which contains the gene and is deposited under the Budapest Treaty on December 1, 2009, is deposited with the American Type Culture Collection Center 'Patent Deposit, 10801 University Boulevard, Manassas, VA 20110-2209, and the ATCC deposit number. PTA-10511. In the previously published Thraustochytrium PUFA synthase system, the PUFA synthase gene corpse/3⁄4/PE4 2 has been clustered and configured to be transcribed. This is also the case from the Thraustochytrium strain ATCC PTA-10212 / with i5 Ke 2 . The concept of discovery/»K2 is 693 nucleotides at the beginning and divergently transcribed. The BAC strain pDS127 (also known as LuMaBAC 9K17) exhibits strong hybridization of the probe to both the DH region and the ER region, as well as DNA sequencing selected for /gene. The BAC strain pDS127, which contains the corpse 3 gene's was deposited with the American Type Culture Collection Center, Patent Depository, 10801 University Boulevard, Manassas, VA 20110-2209, and granted to the ATCC under the Budapest Treaty on December 1, 2009. Deposit number PTA-10510. The sequencing primers of the DNA sequences of the DH region and the ER region determined in Example 4 were designed using standard methods. To determine the DNA sequence of the Thraustochytrium sp. ATCC PTA-10212 gene, subsequent round DNA sequencing (involving subsequent sequencing primers designed by standard methods) was performed to "walk around" the BAC strain. Each nucleotide base pair of a gene is encompassed by a high quality of at least two independent DNA sequencing reactions having at least a minimum aggregated Fryd score of 40 (99.99% confidence level). Table 7 shows the Thraustochytrium strain ATCC ΡΤΑ-1〇212 sequence identification number: 68), sequence identification number: 70), and />like 3 (SEQ ID NO: 72) polynucleotide sequence when previously published The sequence is identical to the sequence from the Schizochytrium strain ΡΤΑ-9695. Identity is determined by the standard of DNA alignment, the scoring matrix "swgapdnamt" in the "AlignX" program of the VectorNTI program. 124 201038734 Table 7: Percent identity of the ke/PE42, and /> ke3 polynucleotide sequences. Comparison of the source of PFA, PF/I2, and PFA? sequences. PFd/{orfA)^% of PFAl Comparison of sex (or/B) to %A of RFA2 (%r Qr/QMPFA3 symmetry Schizochytrium ATCC 20888 55 54 59 Thraustochytrium aureum ATCC 34304 55 53 Unpublished Thraustochytrium Species 23B ATCC 20892 55 57 62 Schizochytrium sp. PTA-9695 55 52 59 Table 8 shows the Thraustochytrium strain ATCC PTA-10212 Pfalp (SEQ ID NO: 69), Pfa2p (SEQ ID NO: 71), and Pfa3p (SEQ ID NO: 73) Identity of the amino acid sequence when compared to the previously published PUFA synthase amino acid sequence and the sequence from Schizochytrium sp. PTA-9695. Identity is by protein alignment. The standard, the scoring matrix in the "AlignX1" program of the VectorNTI program, is determined by blosum62mt2" Table 8: Percent identity comparison with Pfalp, Pfa2p, and Pfa3p amino acid sequences. Comparison of Pfalp, Pfa2p, and Pfa3p columns. (OrfA) Compare Pfa2p with the same *lk (OrfB) is the same as Pfa2p^j%. Pfa3p (OrfC) is the same as % of 卩 kiss. Schizochytrium strain ATCC 20888 62 57 69 Thraustochytrium aureum ATCC 34304 58 54 Unpublished Thraustochytrium strain 23B ATCC 20892 54 54 71 Schizochytrium sp. PTA-9695 59 53 73 Example 6 Performing domain analysis to separately annotate the sequence of Schizochytrium sp. ATCC PTA-10212 PFW, and the area of synthetase and the active position of 125 201038734 The coordinates are identified based on homology to the known PUFA synthase, fatty acid synthase', and polyketide synthase domains. Table 9 shows the associations with Schizochytrium sp. ATCC PTA-10212 corpse E4/ Active position. Table 9: Thraustochytrium strain ATCC PTA-10212 ke/domain analysis domain DNA position ΑΑ position address DNA position ΑΑ position KS Sequence identification number: 68 of 13-1362 Sequence identification number: 69 of 5-545 Active-DXAC» Sequence ID: 68 601~612 Sequence ID: 69 C204 (Sequence ID: 74) (Sequence ID: 75) (Sequence ID: 43) End-GFGG Sequence ID: 68 1351-1362 sequence identification No.: 451454 of 69 (sequence identification number: 44) (sequence identification number: 45) MAT sequence identification number: 68 丨 783-2703 Sequence identification number: 69 of 595-901 active GHSIG (sequence identification number: 46) sequence Identification number: 2083-2085 of 68 (nose identification number: 116) Serial identification number: 69 S695 ACP Sequence identification number: 68 of 3208·6510 (sequence identification number: 78) Serial identification number: 69 of 1070-2170 ( Sequence identification number: 79) ACPI domain sequence identification number: 68 of 3280-3534 (sequence identification number: 80) 109Φ1178 (sequence identification number: 81) ACPI activity LGIDS* (sequence identification number: 47) Sequence identification number: 68 3403-3405 Sequence identification number: 69 S1135 ACP2 domain sequence identification number: 68 of 3607-3861 Sequence identification number: 69 of 1203-1287 (sequence identification number: 83⁄4 (sequence identification number: 83) ACP2 active LGIDS* sequence identification No. · Hu 3371 3732 Sequence identification number: 69 S1244 (sequence identification number: 47) ACP3 domain serial identification number: 68 of 39344185 Serial identification number: 6 132-12-1 of 9 (sequence identification number: 84) (sequence identification number: 85) ACP3 activity LGIDS* sequence identification number: 68, 40,574,059 sequence identification number: 69, S1353 (sequence identification number: 47) ACP4 domain sequence identification number: 68214515 of 68 Sequence identification number: 1421-1505 of 69 (sequence identification number: 86) (sequence identification number: 87) ACP4 activity LGIDS* Sequence identification number: 68 of 43844386 Sequence identification number: 69 of S1462 (sequence identification number: 47) 126 201038734 Ο ο Domain DNA position ΑΑ Location address DNA position AA position ACP5 domain Sequence identification number: 68 4589*4842 Sequence identification number: 69 of 1530-1614 (sequence identification number: 88) (sequence identification number: 89 ACP5 activity LGIDS* Sequence identification number: 4711771313 of sequence identification number: 69 S1571 f戽 column identification number: 47) ACP6 domain sequence identification number: 68 of 4915-5169 Sequence identification number: 69 of 1639-1723 (sequence) Identification number: 90) (sequence identification number: 91) ACP6 active LGIDS* Serial identification number: 68 of 5 038-5040 Sequence identification number: S1680 of 69 (column identification number: 47) ACP7 domain sequence identification number: 5242-54% of 68 Sequence identification number: 69 of 1748-1832 (sequence identification number: 92) (sequence identification number :93) ACP7 activity LGIDS* Sequence identification number: 68 of 5365-5367 Sequence identification number: 69 of S1789 (column identification number: 47) ACP8 domain sequence identification number: 68 of 5569-5823 (sequence identification number: 94) sequence Identification number: 1857-1941 of 69 (sequence identification number: 95) ACP8 activity LGIDS* Sequence identification number: 6692-5694 of sequence identification number: 69 of S1898 (sequence identification number: 47) ACP9 domain sequence identification number: 68 58964150 Sequence identification number: 1966-2050 of 69 (sequence identification number: 96) (sequence identification number: 97) ACP9 activity LGIDS* sequence identification number: 68 of 6019^6021 sequence identification number: 69 of S2007 (sequence identification number: 47 ) ACP10 domain sequence identification number: 68 of 6199~6453 Sequence identification number: 69 of 2067-2151 (sequence identification number: 98) (sequence Identification number: 99) ACT10 active LGIDS * Identification number sequence: SEQ ID. No 68 63224324: 69 S2108 (SEQ ID. No: 47) " core region "

KR 序列辨識编號：68 的 6808-8958 (序列辨識編號： 100) 序列辨識編號：69 的 2270-2986 (序列辨識編號： 101) 序列辨識編號：68 的 7198·8]00 (序列辨識編號： 116) 序列辨識编號：69 的 2400-2600 (序列辨識編號： II： 序列辨識編號：68 序列辨識编號：69 LxxtfccxGxxxxP 序列辨識編號：68 序列辨識编號： DH 的 8203-8241 的 2735-2747 的 8203-8241 的 2735-2747 要素 (序列辨識編號： (序列辨識编號： (序列辨識編號： (序列辨識編號 118) 119) (序列辨識編號:50) 118) 119) 127 201038734 破囊壺菌種ATCC PTA-10212 Pfal内的第一個領域為 KS領域。含有編碼破囊壺菌種ATCC PTA-10212 Pfal KS領 域的序列之核苷酸序列於本文中係表示為序列辨識編號： 74，其係對應於序列辨識編號：68的位置13-1362。包含破 囊壺菌種ATCC PTA-10212 Pfal KS領域的胺基酸序列於本 文中係表示為序列辨識編號：75，其係對應於序列辨識編 號：69的位置5-454。KS領域含有一活性位置要素：DXAC* (序列辨識編號：43)，具有對應於序列辨識編號：69的C204 之一 *醯基結合位址。而且，一特性要素係存在於KS領域的 終端：GFGG(序列辨識編號：44)，其係對應於序列辨識編 號：69的位置451-454以及序列辨識編號：75的位置447-450。 破囊壺菌種ATCC PTA-l〇2l2 Pfal内的第二個領域為 MAT領域。含有編碼破囊壺菌種ATCC PTA-10212 Pfal MAT領域的序列之核苷酸序列於本文中係表示為序列辨識 編號：76,其係對應於序列辨識編號：68的位置1783-2703。 包含破囊壺菌種ATCC PTA-10212 Pfal MAT領域的胺基酸 序列於本文中係表示為序列辨識編號：77，其係對應於序 列辨識編號：69的位置595-901。MAT領域含有一活性位置 要素：GHS*XG(序列辨識編號：46)，具有對應於序列辨識 編號：69的S695之一 *醯基結合位址。 破囊壺菌種ATCC PTA-10212 Pfalp的第3個至第丨2個 領域為10個串聯ACP領域，於本文中亦稱為ACPI、ACP2、 ACP3、ACP4 ' ACP5、ACP6、ACP7、ACP8、ACP9，以 及ACP10 〇含有第一個ACP領域，ACPI的核苷酸序列於本 128 201038734 文中係表示為序列辨識編號：80，以及包含序列辨識編號： 68的大約位置3280延伸到大約位置3534的核苷酸序列。含 有ACPI的胺基酸序列’本文中表示為序顺識編號：81， 係包含序列辨識編號：69的大約位置1094延伸到大約位置 1178的胺基酸序列。含有ACP2的核苷酸序列，本文中表示 為序列辨識編號：82，係包含序列辨識編號：砧的大約位 置3607延伸到大約位置3861的核苷酸序列。含有ACP2的胺 基酸序列，本文中表示為序列辨識編號：83 ,係包含序列 辨識編號：69的大約位置1203延伸到大約位置1287的胺基 酸序列。含有ACP3的核苷酸序列，本文中表示為序列辨識 編號：84，係包含序列辨識編號：68的大約位置3934延伸 到大約位置4185的核苷酸序列。含有acp3的胺基酸序列， 本文中表示為序列辨識編號：85 ’係包含序列辨識編號： 69的大約位置1312延伸到大約位置1396的胺基酸序列。含 有ACP4的核苷酸序列，本文中表示為序列辨識編號：86， 係包含序列辨識編號：68的大約位置4261延伸到大約位置 4515的核苷酸序列。含有ACP4的胺基酸序列，本文中表示 為序列辨識編號：87，係包含序列辨識編號：69的大約位 置1421延伸到大約位置1505的胺基酸序列。含有ACP5的核 苷酸序列’本文中表示為序列辨識編號：88，係包含序列 辨識編號：68的大約位置4589延伸到大約位置4842的核苷 酸序列。含有ACP5的胺基酸序列，本文中表示為序列辨識 編號：89，係包含序列辨識編號：69的大約位置1530延伸 到大約位置1614的胺基酸序列。含有ACP6的核苷酸序列， 129 201038734 本文中表不為序列辨識編號：90，係包含序列辨識編號： 68的大@#：置4915延伸到大約位置5169的核|酸序列。含 有ACP6的胺基酸序列，本文中表示為序列辨識編號：91， 係包含序列辨識編號：69的大約位置1639延伸到大約位置 1723的胺基酸序列。含有Acp7的核苷酸序列，本文中表示 為序列辨識編號：92，係包含序列辨識編號：68的大約位 置5242延伸到大約位置5496的核苷酸序列。含有ACP7的胺 基酸序列，本文中表示為序列辨識編號：93 ’係包含序列 辨識編號：69的大約位置1748延伸到大約位置1832的胺基 酸序列。含有ACP8的核苷酸序列，本文中表示為序列辨識 編號：94 ’係包含序列辨識編號：68的大約位置5569延伸 到大約位置5832的核苷酸序列。含有ACP8的胺基酸序列， 本文中表示為序列辨識編號：95，係包含序列辨識編號： 69的大約位置1857延伸到大約位置1941的胺基酸序列。含 有ACP9的核苷酸序列，本文中表示為序列辨識編號：96， 係包含序列辨識編號：68的大約位置5896延伸到大約位置 6150的核苷酸序列。含有ACP9的胺基酸序列，本文中表示 為序列辨識編號：97，係包含序列辨識編號：69的大約位 置1966延伸到大約位置2050的胺基酸序列。含有ACP10的 核苷酸序列，本文中表示為序列辨識編號：98，係包含序 列辨識編號：68的大約位置6199延伸到大約位置6453的核 苷酸序列。含有ACP10的胺基酸序列，本文中表示為序列 辨識編號：99，係包含序列辨識編號：69的大約位置2067 延伸到大約位置2151的胺基酸序列。全部的10個ACP領域 130 201038734 一起延伸破囊壺菌種ATCC PTA-10212 Pfal的一區域’從序 列辨識編號：68的大約位置3208至大約位置6510，其係對 應於序列辨識編號：69的大約1070至大約2170的胺基酸位 置。含有全部的10個領域的整個ACP區域之核苷酸序列於 本文中係表示為序列辨識編號：78 ;而含有全部的6個領域 的整個ACP區域之胺基酸序列於本文中係表示為序列辨識 編號：79。序列辨識編號：78内之10個ACP領域之重複的 _ 間隔發現為大概每327個核苷酸(經測量之鄰近活性位置絲 〇 胺酸之間的實際胺基酸數目範圍為101至109個胺基酸）。10 個ACP領域之每一者皆含有一泛硫醇結合要素LGIDS*(序 列辨識編號：47)，其中S*為該泛硫醇結合位置絲胺酸(S)。 ' 該泛硫醇結合位置絲胺酸(S )係位於接近每一 A C P領域序列 ' 中心處。6個ACPD領域之每一者的活性位置絲胺酸殘基的 位置（亦即，該泛硫醇結合位置），關於序列辨識編號：69 的胺基酸序列而言，為：ACPI = S1135，ACP2 = S1244， ❹ ACP3 = S1353，ACP4 = S1462，ACP5 = S157卜 ACP6 = S1680，APC7 = S1789，ACP7 = S1789，ACP8 = S1898， ACP9 = S=2007，以及ACP10 = S2108。 破囊壺菌種ATCC PTA-10212 Pfal内的第13個領域為 KR領域。含有編碼pfai KR領域的序列之核苷酸序列於本 文中係表示為序列辨識編號：1〇〇，其係對應於序列辨識編 號：68的位置6808-8958。包含Pfal KR領域的胺基酸序列 於本文中係表示為序列辨識編號：101，其係對應於序列辨 識編號：69的位置2270-2986。在KR領域中為一核心區域(係 131 201038734 包含於序列辨識編號：116的核苷酸序列，以及序列辨識編 號：117的胺基酸序列），與短鏈醛類-脫氫酶具有同源性(KR 為此家族之一成員）。此核心區域係自序列辨識編號·· 68的 大約位置5998延伸至大約6900，其係對應於序列辨識編 號：69的胺基酸位置2000-2300。 破囊壺菌種ATCC PTA-10212 Pfal内的第14個領域為 DH領域。含有編碼Pfal DH領域的序列之核苷酸序列於本 文中係表示為序列辨識編號：118，其係對應於序列辨識編 號：68的位置7027-7065。包含Pfal DH領域的胺基酸序列 於本文中係表示為序列辨識編號：119，其係對應於序列辨 識編號：69的位置2343-2355。DH領域含有守恆性活性位置 要素（參見，Donadio, S. and Katz., L.，Gene 111(1): 51-60 (1992)): LxxHxxxGxxxxP(序列辨識編號：50)。 表10顯示與裂殖壺菌種ATCC PTA-10212 關聯的 領域與活性位置。KR sequence identification number: 68 of 6808-8958 (sequence identification number: 100) Sequence identification number: 69 of 2270-2986 (sequence identification number: 101) Sequence identification number: 68 of 7198·8] 00 (sequence identification number: 116) Sequence identification number: 69 of 2400-2600 (sequence identification number: II: sequence identification number: 68 sequence identification number: 69 LxxtfccxGxxxxP sequence identification number: 68 sequence identification number: DH of 8203-8241 of 2735-2747 2735-2747 element of 8203-8241 (sequence identification number: (sequence identification number: (sequence identification number: (sequence identification number 118) 119) (sequence identification number: 50) 118) 119) 127 201038734 Thraustochytrium The first domain in the ATCC PTA-10212 Pfal is the KS domain. The nucleotide sequence containing the sequence encoding the Thraustochytrium sp. ATCC PTA-10212 Pfal KS domain is represented herein as the sequence ID: 74, Corresponds to position 13-1362 of sequence identification number: 68. The amino acid sequence comprising the Thraustochytrium sp. ATCC PTA-10212 Pfal KS domain is represented herein as sequence identification number: 75, which corresponds to the sequence Identification number: position 5-454 of the KS field. The KS field contains an active position element: DXAC* (sequence identification number: 43) with one of the C204 corresponding to the sequence identification number: 69 * 醯 结合 binding address. The characteristic element is the terminal existing in the KS field: GFGG (sequence identification number: 44), which corresponds to position 451-454 of sequence identification number: 69 and position 447-450 of sequence identification number: 75. The second domain in the ATCC PTA-l〇2l2 Pfal is the MAT field. The nucleotide sequence containing the sequence encoding the Thraustochytrium sp. ATCC PTA-10212 Pfal MAT is represented herein as the sequence ID: 76. It corresponds to position 1783-2703 of sequence identification number: 68. The amino acid sequence containing the Thraustochytrium sp. ATCC PTA-10212 Pfal MAT is represented herein as sequence identification number: 77, which corresponds to the sequence Identification number: position 59 of 595-901. The MAT field contains an active position element: GHS*XG (sequence identification number: 46), with one of the S695 corresponding to the sequence identification number: 69 * thiol binding address. Pot fungus ATCC PTA-10212 Pfalp The 3rd to the 2nd fields are 10 series ACP fields, also referred to herein as ACPI, ACP2, ACP3, ACP4 'ACP5, ACP6, ACP7, ACP8, ACP9, and ACP10 〇 containing the first ACP field. The nucleotide sequence of ACPI is represented herein as Sequence Identification Number: 80, and a nucleotide sequence comprising sequence identification number: 68 extending from position 3280 to position 3534. The amino acid sequence containing ACPI is shown herein as the sequence number: 81, which comprises an amino acid sequence of sequence identification number: 69 extending from about position 1094 to about position 1178. The nucleotide sequence containing ACP2, designated herein as Sequence ID: 82, contains the sequence number: the approximate position 3607 of the anvil extends to a nucleotide sequence at position 3861. The amino acid sequence containing ACP2, designated herein as Sequence Identification Number: 83, comprises an amino acid sequence extending from about position 1203 of sequence identification number: 69 to about position 1287. The nucleotide sequence containing ACP3, designated herein as Sequence Identification Number: 84, comprises a nucleotide sequence of sequence identification number: 68 extending from position 3934 to position 4185. The amino acid sequence containing acp3, designated herein as the sequence number: 85' contains the amino acid sequence of sequence identification number: 69 extending from position 1312 to position 1396. A nucleotide sequence comprising ACP4, designated herein as Sequence ID: 86, comprises a nucleotide sequence of sequence identification number: 68 extending from approximately position 4261 to approximately position 4515. The amino acid sequence containing ACP4, designated herein as Sequence Identification Number: 87, comprises an amino acid sequence extending from position 1421 of sequence number: 69 to approximately position 1505. The nucleotide sequence containing ACP5' is represented herein as sequence identification number: 88, which comprises a nucleotide sequence of sequence identification number: 68 extending from position 4589 to position 4842. The amino acid sequence containing ACP5, designated herein as Sequence Identification Number: 89, comprises an amino acid sequence extending from about position 1530 of sequence identification number: 69 to about position 1614. Nucleotide sequence containing ACP6, 129 201038734 This is not a sequence identification number: 90, which contains a sequence identification number: 68 large @#: a 4919 extension to a nuclear/acid sequence at position 5169. The amino acid sequence comprising ACP6, designated herein as Sequence Identification Number: 91, comprises an amino acid sequence of sequence identification number: 69 extending from about position 1639 to about position 1723. The nucleotide sequence containing Acp7, designated herein as Sequence Identification Number: 92, comprises a nucleotide sequence comprising the sequence position 5242 of Sequence Identification Number: 68 extending to approximately position 5496. The amino acid sequence containing ACP7, designated herein as the sequence number: 93' contains the amino acid sequence of sequence identification number: 69 extending from approximately position 1748 to approximately position 1832. A nucleotide sequence comprising ACP8, designated herein as a sequence identification number: 94' contains a nucleotide sequence of sequence identification number: 68 from about position 5569 to about position 5832. The amino acid sequence containing ACP8, designated herein as Sequence Identification Number: 95, comprises an amino acid sequence extending from about position 1857 of sequence identification number: 69 to about position 1941. A nucleotide sequence comprising ACP9, designated herein as Sequence Identification Number: 96, comprises a nucleotide sequence of sequence identification number: 68 extending from position 5896 to approximately position 6150. The amino acid sequence containing ACP9, designated herein as Sequence Identification Number: 97, contains an amino acid sequence of sequence position number 1966 extending to approximately position 2050. The nucleotide sequence containing ACP10, designated herein as Sequence ID: 98, contains a nucleotide sequence of sequence identification number: 68 extending from position 6199 to position 6453. The amino acid sequence containing ACP10, designated herein as Sequence Identification Number: 99, contains the amino acid sequence of sequence identification number: 69, approximately position 2067, extending to approximately position 2151. All 10 ACP domains 130 201038734 together extend a region of the Thraustochytrium ATCC PTA-10212 Pfal 'from the sequence identification number: 68 to the approximate position 3208 to approximately position 6510, which corresponds to the sequence identification number: 69 approximately Amino acid position from 1070 to about 2170. The nucleotide sequence of the entire ACP region containing all 10 domains is represented herein as sequence identification number: 78; and the amino acid sequence containing the entire ACP region of all 6 domains is represented herein as a sequence. Identification number: 79. Sequence identification number: Repeated _ intervals in 10 ACP domains within 78 were found to be approximately every 327 nucleotides (the actual number of amino acids between the measured active sites of proline was between 101 and 109) Amino acid). Each of the 10 ACP domains contains a panthenol-binding element LGIDS* (serial identification number: 47), where S* is the panthenol-binding site serine (S). The panthenol-binding position of the serine acid (S) line is located near the center of each A C P domain sequence. The position of the active position of the serine residue in each of the six ACPD domains (ie, the panthenol binding site), for the amino acid sequence of sequence number: 69, is: ACPI = S1135, ACP2 = S1244, ❹ ACP3 = S1353, ACP4 = S1462, ACP5 = S157, ACP6 = S1680, APC7 = S1789, ACP7 = S1789, ACP8 = S1898, ACP9 = S=2007, and ACP10 = S2108. The thirteenth field of the Thraustochytrium strain ATCC PTA-10212 Pfal is the KR field. A nucleotide sequence comprising a sequence encoding the pfai KR domain is referred to herein as a sequence number: 1 〇〇, which corresponds to position 6808-8958 of sequence identification number: 68. The amino acid sequence comprising the Pfal KR domain is referred to herein as Sequence Identification Number: 101, which corresponds to position 2270-2986 of Sequence Identification Number: 69. In the KR domain, a core region (line 131 201038734 is included in the nucleotide sequence of sequence identification number: 116, and the amino acid sequence of sequence identification number: 117) has homology to the short-chain aldehyde-dehydrogenase. Sex (KR is a member of this family). This core region extends from approximately position 5998 of the sequence identification number 68 to approximately 6900, which corresponds to the amino acid position of the sequence identification number: 69 from 2000 to 2300. The 14th field of the Thraustochytrium strain ATCC PTA-10212 Pfal is the DH field. A nucleotide sequence comprising a sequence encoding the Pfal DH domain is referred to herein as Sequence Identification Number: 118, which corresponds to position 7027-7065 of Sequence Identification Number: 68. The amino acid sequence comprising the Pfal DH domain is represented herein as Sequence Identification Number: 119, which corresponds to position 2343-2355 of Sequence Identification Number: 69. The DH domain contains conserved active positional elements (see, Donadio, S. and Katz., L., Gene 111(1): 51-60 (1992)): LxxHxxxGxxxxP (SEQ ID NO: 50). Table 10 shows the domains and active positions associated with Schizochytrium sp. ATCC PTA-10212.

132 201038734 表10 :破囊壺菌種ΑΤ(ΧΡΤΑ-1021271Ε42領域分析 領域 DNA位置 AA位置 位址 DNA位置 AA位置一 KS 序列辨識編號：70 的 10-1320 序列辨識編號：71 的 Φ440 DXAC* 序列辨識編號：70 的 5^-573 序列辨識編號：71 的 C191 (序列辨戴編號:102) (序列辨識編號:103) (序列辨識編號:43) End-GFGG 序列辨識編號：70 的 1267-1278 序列辨識編號：71 的423426 (序列辨識編號：44) CLF 序列辨識編號：70 的 1378-2700 序列辨識編號：Ή 的460-900 (序列辨識編號:104) (序列辨識編號:105) AT 序列辨識編號：70 的28484200 序列辨識編號：*71 的950>1400 GxS*xG 序列辨識編號：70 的3361-3363 序列辨識編號：71 的 S1121 (序列辨識編號:106) (序列辨識編號:107) (序列辨識編號:52) ER 序列辨識編號：70 的4498-5700 序列辨識編號：Ή 的 1500-1900 (序列辨識編號:1〇8) (序列辨識編號_· 109) ❹ 破囊壺菌種ATCC PTA-10212 Pfa2内的第1個領域為 KS領域。含有編碼破囊壺菌種ATCCPTA-10212Pfa2KS領 域的序列之核苷酸序列於本文中係表示為序列辨識編號： 102，其係對應於序列辨識編號：70的位置10-1320。包含 破囊壺菌種ATCC PTA-10212 Pfa2 KS領域的胺基酸序列於 本文中係表示為序列辨識編號：1〇3，其係對應於序列辨識 編號：71的位置4-440。KS領域含有一活性位置要素： DXAC*(序列辨識編號：43)，具有對應於序列辨識編號： 71的C191之一*醯基結合位址。而且，一特性要素係存在於 KS領域的終端：GFGG(序列辨識編號：44) ’其係對應於序 列辨識編號：71的位置423-426以及序列辨識編號：70的位 置1267-1278 。132 201038734 Table 10: Thraustochytrium species (ΧΡΤΑ-1021271Ε42 field of analysis DNA location AA position address DNA position AA position-KS Sequence identification number: 70 10-1320 Sequence identification number: 71 Φ440 DXAC* Sequence identification No.: 70 of 5^-573 Sequence identification number: 71 C191 (sequence identification number: 102) (sequence identification number: 103) (sequence identification number: 43) End-GFGG sequence identification number: 1267-1278 sequence of 70 Identification number: 423426 of 71 (sequence identification number: 44) CLF sequence identification number: 70 of 1378-2700 Sequence identification number: 460 460-900 (sequence identification number: 104) (sequence identification number: 105) AT sequence identification number 28484200 of :70 Sequence identification number: 950 of *71 > 1400 GxS*xG Sequence identification number: 3361-3363 of 70 Sequence identification number: 71 S1121 (sequence identification number: 106) (sequence identification number: 107) (sequence identification No.: 52) ER Sequence identification number: 70 4498-5700 Sequence identification number: 1500 1500-1900 (sequence identification number: 1〇8) (sequence identification number _· 109) 破 Thraustochytrium species A The first domain in TCC PTA-10212 Pfa2 is the KS domain. The nucleotide sequence containing the sequence encoding the Thraustochytrium sp. ATCCPTA-10212Pfa2KS domain is represented herein as the sequence identification number: 102, which corresponds to the sequence. Identification number: position 10-1320 of 70. The amino acid sequence comprising the Thraustochytrium sp. ATCC PTA-10212 Pfa2 KS domain is represented herein as the sequence identification number: 1〇3, which corresponds to the sequence identification number: Position 71 of the 71. The KS field contains an active positional element: DXAC* (sequence identification number: 43) having one of the C191 corresponding to the sequence identification number: 71; the 醯-based binding address. Terminals present in the KS field: GFGG (Sequence Identification Number: 44) 'This corresponds to position identification number 423-426 of sequence identification number: 71 and position 1267-1278 of sequence identification number: 70.

破囊壺菌種ATCC PTA-10212 Pfa2内的第2個領域為 CLF領域。含有編碼破囊壺菌種ATCC PTA-10212 Pfa2 CLF 133 201038734 領域的序列之核苷酸序列於本文中係表示為序列辨識編 號：104，其係對應於序列辨識編號：70的位置1378-2700。 包含破囊壺菌種ATCC PTA-10212 Pfa2 CLF領域的胺基酸 序列於本文中係表示為序列辨識編號：105，其係對應於序 列辨識編號：71的位置460-900。The second field within the Thraustochytrium strain ATCC PTA-10212 Pfa2 is the CLF field. The nucleotide sequence containing the sequence encoding the Thraustochytrium sp. ATCC PTA-10212 Pfa2 CLF 133 201038734 is represented herein as the sequence ID: 104, which corresponds to position 1378-2700 of sequence identification number: 70. The amino acid sequence comprising the Thraustochytrium sp. ATCC PTA-10212 Pfa2 CLF domain is represented herein as Sequence Identification Number: 105, which corresponds to position 460-900 of Sequence Identification Number: 71.

破囊壺菌種ATCC PTA-10212 Pfa2内的第3個領域為AT 領域。含有編碼破囊壺菌種ATCC PTA-10212 Pfa2 AT領域 的序列之核苷酸序列於本文中係表示為序列辨識編號： 106，其係對應於序列辨識編號：70的位置2848-4200。包 含破囊壺菌種ATCC PTA-10212 Pfa2 AT領域的胺基酸序列 於本文中係表示為序列辨識編號：107，其係對應於序列辨 識編號：71的位置950-1400。AT領域含有一活性位置要素 GxS*xG(序列辨識編號：50)，其為醯基轉移酶(AT)蛋白質 的特徵，具有對應於序列辨識編號：71的位置S1121的一活 性位置絲胺酸殘基。 破囊壺菌種ATCC PTA-10212 Pfa2内的第4個領域為 ER領域。含有編碼破囊壺菌種ATCC PTA-10212 Pfa2ER領 域的序列之核苷酸序列於本文中係表示為序列辨識編號： 108，其係對應於序列辨識編號：70的位置4498-5700。包 含破囊壺菌種ATCC PTA-10212 Pfa2 ER領域的胺基酸序列 於本文中係表示為序列辨識編號：109，其係對應於序列辨 識編號：71的位置1500-1900。 表11顯示與裂殖壺菌種ATCC ΡΤΑ-10212 關聯的 領域與活性位置。 201038734 表11 :破囊壺菌種ATCCPTA-l〇2l2尸柯3領域分析 領域 DNA位置 AA位置 位址 DNA位置 ΑΑ位置 DH1 序列辨識編號：72 的 1-1350 序列辨識編號：73 的M50 FxxH*F 序列辨識編號：72 的934·936 序列辨識編號：乃 的 Η312 (序列辨識編號:110) (序列辨識編號:ill) (序列辨識編號:53) DH2 序列辨識編號：72 的 1501-2700 序列辨識編號=73 的501-900 FxxH*F 序列辨識編號：72 的2401-2403 序列辨識編號：« 的 Η801 (序列辨書鱗號:112) (序列辨識编號:113) (序列辨識編號:53) ER 序列辨識編號：72 ^28484212 序列辨識編號：73 的 95W404 (序列辨識編號:114) (序列辨識編號:II5) 破囊壺菌種ATCC PTA-10212 Pfa3内的第一與第二個 領域為DH領域，於本文中各別地稱為DH1和DH2。含有編 碼破囊壺菌種ATCC PTA-10212 Pfa3 DH1領域的序列之核 苷酸序列於本文中係表示為序列辨識編號：11〇，其係對應 於序列辨識編號：72的位置1-1350。包含破囊壺菌種ATCC PTA-10212 Pfa3 DH1領域的胺基酸序列於本文中係表示為 序列辨識編號：111，其係對應於序列辨識編號：73的位置 1-450。含有編碼破囊壺菌種ATCC PTA-10212 Pfa3 DH2領 域的序列之核苷酸序列於本文中係表示為序列辨識編號： 112，其係對應於序列辨識編號：72的位置1501_27〇〇。包 含破囊壺菌種ATCC PTA-10212 Pfa3 DH2領域的胺基酸序 列於本文中係表示為序列辨識編號：113，其係對應於序列 辨識編號：73的位置501 -900。該等DH領域含有一活性位置 要素：FxxH*F(序列辨識編號：53)。含有DH1之中的活性 位置要素之核苷酸序列係對應於序列辨識編號：72的位置 934-936，而含有DH2之中的活性位置要素之核苷酸序列係 135 201038734 對應於序列辨識編號：72的位置2401-2403。於該要素 FxxH*F中的該活性位置H*係根據來自Leesong等人， Structure 4:253-64 (1996)及Kimber 等人，J Biol Chem. 279:52593-602 (2004)的資料，DH1中的該活性位置Η*對應 於序列辨識編號：73的Η312以及DH2中的該活性位置Η*對 應於序列辨識編號：73的Η801。 破囊壺菌種ATCC PTA-10212 Pfa3内的第3個領域為 ER領域。含有編碼破囊壺菌種ATCC PTA-10212 Pfa3 ER領 域的序列之核苷酸序列於本文中係表示為序列辨識編號： 114，其係對應於序列辨識編號：72的位置2848-4200。包 含破囊壺菌種ATCC PTA-10212 Pfa3 ER領域的胺基酸序列 於本文中係表示為序列辨識編號：115，其係對應於序列辨 識編號：73的位置950-1400。 實施例7 於裂殖壺菌種ATCC 20888内不活化天然的PUFA合成 酶基因’以產生PUFA營養缺陷型，以及用外源引入的同源 性基因取代此等不活化的基因以恢復PUFA合成，此點先前 已經經證明且被說明。參見，例如，美國專利案第7,217,856 號，以其之整體併入本文中作為參考資料。來自裂殖壺菌 種ATCC 20888之3種PUFA合成酶基因先前已經稱為orfA、 orfB，和orfC，各別地對應於本文中使用的朽ay、， 以及/7^3的命名。同前。 於裂殖壺菌種ATCC 20888内之天然的orfA基因係藉由 同源性重組予以取代，接著用一載體予以轉形，該載體含 136 201038734 有被來自orfA的側邊區域的序列所圍繞之抗吉歐徽素 (Zeocin)TM標誌。產生了缺少功能性〇rfA基因的一突變菌 株。該突變菌株為營養缺陷型的且需要用於生長之PUFA補 充。 裂殖壺菌種ATCC PTA-9695 (序列辨識編號：1) 係被選殖至表現載體PREZ37之内以產生pREZ345。該表現 載體含有來自裂殖壺菌種ATCC 20888的天然的orfA基因位 置的側邊區域之大概2 kb的DNA。缺少功能性orfA的裂殖 壺菌種ATCC 20888突變體係用含有的pREZ345、經由 帶有酵素預處理的電穿孔予以轉形（參見以下）。根據在該突 變體内的抗吉歐黴素™標誌之側邊以及在PREZ345内的 基因的側邊之同源的區域，發生雙交換重組，藉此 P/M Y係插入至天然的orfA位置中。裂殖壺菌種ATCC PTA-9695 (序列辨識編號：1)之重組作用恢復了在缺 少orfA之裂殖壺菌種ATCC 20888突變體内之PUFA的生 產。簡言之，令細胞於M2B液體培養基（參見下列段落）中伴 隨200 rpm震盪在30。(：下生長歷時3天。收穫細胞且使用標 準的技術將該等脂肪酸轉變成甲酯。脂肪酸形態係使用附 有火焰離子偵測器之氣相層析(GC-FID)決定為脂肪酸曱酯 (FAME)。含有功能性orfA基因之天然的裂殖壺菌種ATcc 20888菌株生產2.3 : 1的比率之DHA與DPA n-6。含有代替 不活化的orfA基因之裂殖壺菌種ATCC PTA-9695户冗47 (序 列辨識編號：1)的重組型菌株亦生產2.4 : 1的比率之DHA 與DPA n-6。該重組型菌株的epa含量為2.7%的脂肪酸甲酯 137 201038734 (FAME)，DPAn-3含量為0.7%，DPAn-6含量為8.8%，以及 DHA含量為21.2%。 M2B培養基-10 g/L葡萄糖、0.8 g/L (NH4)2S04、5 g/L Na2S04、2 g/L MgS04.7H20、0.5 g/L KH2P〇4、0.5 g/L Κα、 0· 1 g/L CaCl2.2H20、0· 1 M MES (pH 6.0) 0· 1 % PB26金屬， 以及0·1% PB26維生素(v/v)。PB26維生素由50 mg/mL維生 素B12、100 pg/mL硫胺素，以及100 pg/mL Ca-泛酸所組 成。將？826金屬調整至卩^14.5，以及由3 8/1^6804.7«；20、 1 g/L MnCl2.4H20、800 mg/mL ZnS04.7H20、20 mg/mL CoCl2*6H2〇、10 mg/mL Na2Mo〇4*2H2〇、600 mg/mL CuS04.5H20 ’ 以及800 mg/mLNiS04.6H20所組成。將PB26 儲備液分別地經過濾器滅菌以及在高壓蒸氣滅菌之後添加 至肉湯。葡萄糖、ΚΗ2Ρ04，以及CaCl2.2H20係在混合之前 與肉湯成分剩餘物分別地各自高壓蒸氣滅菌以避免鹽類沉 澱並使碳水化合物焦糖化。全部的培養基成分係購自於 Sigma Chemical (St. Louis, MO) ° 帶有酵素預處理的電穿孔-細胞係以200 rpm於一震盪 器上於50 mL的M50-20培養基（參見美國公開案第號 2008/0022422)中、30°C下生長歷時2天。該等細胞係以1 : 100予以稀釋至M2B培養基之内並生長過夜（16-24 h)，企圖 達到對數中期相的生長（1.5-2.5的OD600)。該等細胞係以大 約3000 X g於50 mL圓錐形管中予以離心歷時5 min。移除懸 浮液’以及將細胞再懸浮於適當體積的pH 5.5之1 Μ甘露糖 醇之中，以達到2 OD600單位的最終濃度。將5 mL的細胞 138 201038734 分量至25 mL震盪燒瓶内以及用10 mM CaCl2 (經過濾器滅 菌的1.0 Μ儲備液)和0.25 mg/mL蛋白酶XIV(經過渡器滅 ij 的 10 mg/mL 儲備液；sigma-Aldrich, St. Louis, MO)予以The third domain within the Thraustochytrium strain ATCC PTA-10212 Pfa2 is the AT field. The nucleotide sequence containing the sequence encoding the Thraustochytrium sp. ATCC PTA-10212 Pfa2 AT domain is represented herein as Sequence Identification Number: 106, which corresponds to position 2848-4200 of Sequence Identification Number:70. The amino acid sequence comprising the Thraustochytrium sp. ATCC PTA-10212 Pfa2 AT domain is represented herein as Sequence Identification Number: 107, which corresponds to position 950-1400 of Sequence Identification Number: 71. The AT domain contains an active positional element GxS*xG (SEQ ID NO: 50) which is characteristic of a thiotransferase (AT) protein having an active position of a serine residue corresponding to position S1121 of sequence identification number: 71. base. The fourth field within the Thraustochytrium strain ATCC PTA-10212 Pfa2 is the ER field. The nucleotide sequence containing the sequence encoding the Thraustochytrium sp. ATCC PTA-10212 Pfa2ER is represented herein as the sequence ID: 108, which corresponds to position 4498-5700 of sequence identification number: 70. The amino acid sequence comprising the Thraustochytrium sp. ATCC PTA-10212 Pfa2 ER domain is represented herein as Sequence Identification Number: 109, which corresponds to the position of Sequence Identification Number: 71, 1500-1900. Table 11 shows the domains and active positions associated with Schizochytrium sp. ATCC ΡΤΑ-10212. 201038734 Table 11: Thraustochytrium species ATCCPTA-l〇2l2 corpse 3 field analysis field DNA position AA position address DNA position ΑΑ position DH1 Sequence identification number: 72 1-1350 Sequence identification number: 73 of M50 FxxH*F Sequence identification number: 934·936 of 72. Sequence identification number: Η 312 (sequence identification number: 110) (sequence identification number: ill) (sequence identification number: 53) DH2 sequence identification number: 72 1501-2700 sequence identification number 501-900 FxxH*F of =73 Sequence identification number: 2401-2403 of 72 Sequence identification number: « Η 801 (sequence identification number: 112) (sequence identification number: 113) (sequence identification number: 53) ER Sequence identification number: 72 ^28484212 Sequence identification number: 73 of 95W404 (sequence identification number: 114) (sequence identification number: II5) Thraustochytrium sp. ATCC PTA-10212 The first and second fields in Pfa3 are DH fields , referred to herein as DH1 and DH2, respectively. The nucleotide sequence containing the sequence encoding the Thraustochytrium sp. ATCC PTA-10212 Pfa3 DH1 domain is represented herein as the sequence identification number: 11 〇, which corresponds to position 1-1350 of sequence identification number: 72. The amino acid sequence comprising the Thraustochytrium sp. ATCC PTA-10212 Pfa3 DH1 domain is represented herein as Sequence Identification Number: 111, which corresponds to position 1-450 of Sequence Identification Number: 73. The nucleotide sequence containing the sequence encoding the Thraustochytrium sp. ATCC PTA-10212 Pfa3 DH2 is represented herein as sequence identification number: 112, which corresponds to position 1501_27 of sequence identification number: 72. The amino acid sequence comprising the Thraustochytrium sp. ATCC PTA-10212 Pfa3 DH2 is listed herein as Sequence Identification Number: 113, which corresponds to position 501-900 of Sequence Identification Number: 73. These DH fields contain an active positional element: FxxH*F (SEQ ID NO: 53). The nucleotide sequence containing the active position element in DH1 corresponds to position 934-936 of sequence identification number: 72, and the nucleotide sequence 135 201038734 containing the active position element among DH2 corresponds to the sequence identification number: The position of 72 is 2401-2403. The active position H* in the element FxxH*F is based on data from Leesong et al., Structure 4: 253-64 (1996) and Kimber et al., J Biol Chem. 279: 52593-602 (2004), DH1. The active position Η* corresponding to Η 312 of sequence identification number: 73 and the active position Η* in DH2 correspond to Η 801 of sequence identification number: 73. The third field in the genus Thraustochytrium ATCC PTA-10212 Pfa3 is the ER field. The nucleotide sequence containing the sequence encoding the Thraustochytrium sp. ATCC PTA-10212 Pfa3 ER domain is represented herein as the sequence number: 114, which corresponds to position 2848-4200 of sequence number: 72. The amino acid sequence comprising the Thraustochytrium sp. ATCC PTA-10212 Pfa3 ER domain is represented herein as Sequence Identification Number: 115, which corresponds to position 950-1400 of Sequence Identification Number: 73. Example 7 does not activate the native PUFA synthase gene in the Schizochytrium sp. ATCC 20888 to produce PUFA auxotrophy, and replaces these inactivated genes with exogenously introduced homologous genes to restore PUFA synthesis, This point has been previously proven and explained. See, for example, U.S. Patent No. 7,217,856, incorporated herein by reference in its entirety. The three PUFA synthase genes from Schizochytrium sp. ATCC 20888 have previously been referred to as orfA, orfB, and orfC, each corresponding to the nomenclature used herein, and the nomenclature of /7^3. Cit. The native orfA gene in Schizochytrium sp. ATCC 20888 is replaced by homologous recombination followed by transformation with a vector containing 136 201038734 surrounded by sequences from the side regions of orfA. Anti-Geoic ZeocinTM logo. A mutant strain lacking the functional 〇rfA gene was generated. The mutant strain is auxotrophic and requires PUFA supplementation for growth. The Schizochytrium strain ATCC PTA-9695 (SEQ ID NO: 1) was cloned into the expression vector PREZ37 to generate pREZ345. The expression vector contains approximately 2 kb of DNA from the flanking region of the native orfA gene locus of Schizochytrium sp. ATCC 20888. The Schizochytrium sp. ATCC 20888 mutant system lacking functional orfA was transformed with pREZ345 containing electroporation with enzyme pretreatment (see below). Double-crossover recombination occurs according to the side of the anti-GiomycinTM marker in the mutant and the homologous region of the gene in the PREZ345, whereby the P/MY line is inserted into the native orfA position. . The recombination of Schizochytrium sp. ATCC PTA-9695 (SEQ ID NO: 1) restored the production of PUFA in the absence of orfA of the Schizochytrium sp. ATCC 20888 mutant. Briefly, cells were shaken at 30 rpm in M2B liquid medium (see paragraph below) at 30 rpm. (The growth was carried out for 3 days. The cells were harvested and converted to methyl esters using standard techniques. The fatty acid morphology was determined by gas chromatography with a flame ion detector (GC-FID) as the fatty acid oxime ester. (FAME). The natural Schizochytrium strain ATcc 20888 strain containing the functional orfA gene produces a ratio of DHA and DPA n-6 of 2.3: 1. Contains the Schizochytrium sp. ATCC PTA- instead of the inactivated orfA gene. 9695 households with 47 (sequence identification number: 1) recombinant strains also produced DHA and DPA n-6 in a ratio of 2.4: 1. The recombinant strain had an epa content of 2.7% fatty acid methyl ester 137 201038734 (FAME), The DPAn-3 content was 0.7%, the DPAn-6 content was 8.8%, and the DHA content was 21.2%. M2B medium-10 g/L glucose, 0.8 g/L (NH4) 2S04, 5 g/L Na2S04, 2 g/ L MgS04.7H20, 0.5 g/L KH2P〇4, 0.5 g/L Κα, 0·1 g/L CaCl2.2H20, 0· 1 M MES (pH 6.0) 0· 1 % PB26 metal, and 0.1% PB26 vitamin (v/v). PB26 vitamin consists of 50 mg/mL vitamin B12, 100 pg/mL thiamine, and 100 pg/mL Ca-pantothenic acid. Adjust 826 metal to 卩^14.5, and by 3 8/1^6 804.7«; 20, 1 g/L MnCl2.4H20, 800 mg/mL ZnS04.7H20, 20 mg/mL CoCl2*6H2〇, 10 mg/mL Na2Mo〇4*2H2〇, 600 mg/mL CuS04.5H20 ' and 800 mg/mL NiS04.6H20. The PB26 stock solution was separately sterilized by filter and added to the broth after autoclaving. Glucose, ΚΗ2Ρ04, and CaCl2.2H20 were separately mixed with the broth residue before mixing. Autoclaved to avoid salt precipitation and caramelization of carbohydrates. All media components were purchased from Sigma Chemical (St. Louis, MO) ° Electroporation-cell line with enzyme pretreatment at 200 rpm The cells were grown in 50 mL of M50-20 medium (see US Publication No. 2008/0022422) for 2 days at 30 ° C. The cell lines were diluted 1:100 into M2B medium and grown overnight. (16-24 h), attempting to achieve growth in the mid-log phase (OD600 of 1.5-2.5). The cell lines were centrifuged in a 50 mL conical tube for approximately 5 min at approximately 3000 X g. The suspension was removed and the cells were resuspended in an appropriate volume of 1 mM mannitol at pH 5.5 to achieve a final concentration of 2 OD600 units. 5 mL of cell 138 201038734 fraction into a 25 mL shake flask and 10 mM CaCl2 (filter sterilized 1.0 Μ stock solution) and 0.25 mg/mL protease XIV (10 mg/mL stock solution via a transition annihilator); sigma-Aldrich, St. Louis, MO)

修改。燒瓶係於3〇t及大約100 rpm於一震盪器上孵育歷時 4h。於顯微鏡下監看細胞以決定原生質體化的程度，帶有 所欲的單一細胞。令細胞於圓底管（亦即，l4inLFaiconTM 管，BD Biosciences，San Jose，CA)内以大約2500 xg予以 離心歷時5 min。移除懸浮液，以及該等細胞係用5 mL冰冷 的10%甘油予以溫和地再懸浮。令該等細胞於圓底管内以 大約2500 X g予以再次離心歷時5 min。移除懸浮液，以及 該等細胞係用500 pL冰冷的10%甘油使用寬孔的微量吸管 予以溫和地再懸浮。將9〇 pL的細胞分量至預冷的電_光析 管（Gene Pulser®光析管_〇.1 cm間隙或〇.2 cm間隙， Bio-Rad，Hercules，CA)中。將1 至5 pg的DNA(以低於 或等於10 pL體積）添加至光析管，用一微量吸管予以溫和 地混合’以及放置於冰上歷時5 min。細胞係以200 〇hmS (電 阻）、25 pF (電容），以及250V (用於0.1 cm間隙）或5〇〇v(〇.2 cm間隙）予以電穿孔。將0.5 mL的M50-20培養基立即添加至 光析管。接而將細胞轉移至於25 mL震盪燒瓶中之4.5 mL的 M50-20培養基内以及於3〇°C及大約100 rpm於一震盈器上 孵育歷時2-3 h。令該等細胞於圓底管内以大約2500 X g予以 離心歷時5 m i η。移除懸浮液以及將細胞沉澱丸再懸浮於〇. 5 mL的Μ50-20培養基之中。將細胞平盤培養魚合適數目（2至 5)的M2B平盤上，伴隨合適的挑選以及於3〇°C下孵育。 139 201038734 缺少功能性orfA的裂殖壺菌種ATCC 2〇888突變體也用 含有PFW的PREZ345予以轉形，藉此户柯7係隨機與該突變 體結合在一起以及恢復PUFA的生產。 實施例8 破囊壺菌種ATCC ?丁八-1〇212/^7 (序列辨識編號：68) 係被再合成(DNA2.0)且被密碼子最佳化用於裂殖壺菌内表 現(序列辨識編號：120) ’以及被選殖至一表現載體之内來 產生pLR95。密碼子最佳化係使用第22圖中的裂殖壺菌密碼 子使用表來產生。該表現載體含有來自裂殖壺菌種ATCC 20888的天然的orfA基因位置之側邊區域的大概2 kb的 DNA。 來自實施例7之缺少功能性orfA的裂殖壺菌種ATCC 20888突變體係用含有密碼子最佳化的破囊壺菌種ATCC ΡΤΑ-10212ΡΛ4/(序列辨識編號：120)之pLR95、經由帶有 酵素預處理的電穿孔予以轉形（參見實施例7)。根據在該突 變體内的抗吉歐黴素TM標誌之側邊以及在pLR95内的户柯7 基因的側邊之同源的區域，發生雙交換重組，藉此密碼子 最佳化的破囊壺菌種ATCC ΡΤΑ-1〇212户柯/係插入至天然 的orfA位置中。具有密碼子最佳化的破囊壺菌種ATCC PTA-10212尸E4J(序列辨識編號：120)之重組作用恢復了在 缺少orfA之裂殖壺菌種ATCC 20888突變體内之PUFA的生 產。細胞係如實施例7中說明的方式生長且分析FAMEs。含 有功能性orfA基因之天然的裂殖壺菌種ATCC 20888菌株生 產比率25 : 1之DHA和EPA。含有代替不活化的orfA基因之 140 201038734 密碼子最佳化的破囊壺菌種ATCC ΡΤΑ-10212 序列辨 識編號：120)的該重組型菌株生產比率5.4 : 1之DHA和 EPA，進一步展現出裂殖壺菌的PUFA形態可以藉由本文中 說明的核酸分子予以改變。該重組型菌株的EPA含量為 4.4%的FAME，DPAn-3含量為2.3%，DPAn-6含量為 4.9%， 以及DHA含量為24.0%。 缺少功能性orfA的裂殖壺菌種ATCC 20888突變體也用 含有的pLR95予以轉形，藉此係隨機與該突變體 結合在一起以及恢復PUFA的生產。 實施例9 裂殖壺菌種ATCC 20888内的天然的orfB基因係藉由同 源性重組予以取代，接著用一載體經由帶有酵素預處理的 電穿孔予以轉形，該載體含有被來自orffl的側邊區域的序 列所圍繞之抗吉歐黴素™標誌。產生缺少功能性orfB基因 的一突變菌株。該突變菌株為營養缺陷型的且需要用於生 長之PUFA補充。 裂殖壺菌種ATCC PTA-9695尸/^ (序列辨識編號：3) 係被選殖至表現載體pDS 04之内以產生pREZ3 31。該表現載 體含有來自裂殖壺菌種ATCC 20888的天然的orfB基因位置 之側邊區域的大概2 kb的DNA。 缺少功能性orfB之裂殖壺菌種ATCC 20888突變體係用 含有PFA2的pREZ331予以轉形。基於該突變體内的隨機結 合，PUFA生產係藉由裂殖壺菌種ATCC PTA-9695戶柯2 (序 列辨識編號：3)而恢復。細胞係如實施例7中說明的方式生 141 201038734 長且分析FAMEs。含有功能性orfB基因之天然的裂殖壺菌 種ATCC 20888菌株生產2.3 : 1的比率之dha和DPA n-6。含 有取代不活化的orfB基因之裂殖壺菌種ATCC PTA_9695 (序列辨識編號：3)的重組型菌株生產3.5 : 1的比率之 DHA和DPA n-6。該重組型菌株的ΕΡΑ含量為〇 8〇/〇的 FAME，DPA η-3含量為0.1% ’ DPA n-6含量為7.1%，以及 DHA含量為25.1%。 缺少功能性orfB的裂殖壺菌種ATCC 20888突變體也用 含有PFW的pREZ331予以轉形，藉此係插入至天然的 orfB位置中以及恢復PUFA的生產。 實施例10 破囊壺菌種ATCC PTA-10212 PFA2 (序列辨識編號：70) 係被再合成(DNA2.0)且密碼子最佳化為了於裂殖壺菌内表 現(序列辨識編號：121)，以及被選殖至一表現载體之内以 產生PLR85。密碼子最佳化係使用第22圖中的裂殖壺菌密碼 子使用表來產生。該表現載體含有來自裂殖壺菌種ATCC 20888的天然的orfB基因位置之側邊區域的大概2 kb的 DNA。 也於具有改良的DHA生產力之後代菌株的裂殖壺菌種 ATCC 20888中研究orf基因的取代。後代菌株内之天然的 orfB基因係藉由同源性重組予以取代，接著用一載體經由 帶有酵素預處理的電穿孔予以轉形（參見實施例7)，該載體 含有被來自orffl的側邊區域的序列所圍繞之抗吉歐黴素TM 標誌。產生了缺少功能性orfB基因的一突變菌株。該突變 142 201038734 菌株為營養缺陷型的且需要用於生長之PUFA補充。該突變 菌株係用pLR85經由帶有酵素預處理的電穿孔予以轉形（參 見實施例8)，該載體含有密碼子最佳化的破囊壺菌種ATCC PTA-10212 PE42 (序列辨識編號：121)。根據在該突變體内 的抗吉歐黴素TM標誌之側邊以及在pLR85内的PM2基因的 側邊之同源的區域，發生雙交換重組，藉此密碼子最佳化 的破囊壺菌種ATCCPTA-l〇212尸序列辨識編號：121) 係插入至該突變菌株之天然的orfB位置中。具有密碼子最 佳化的破囊壺菌種ATCC ΡΤΑ-10212 序列辨識編 號：121)之重組作用恢復了在缺少orfB之後代菌株突變體内 之PUFA的生產。細胞係如實施例7中說明的方式來生長且 分析FAMEs。該重組型菌株的EPA含量為1.0%的FAME， DPAn-3含量為0.3%，DPAn-6含量為7.0%，以及DHA含量 為 31.0%。 於一待執行的實驗中，來自實施例9之缺少功能性orfB 的裂殖壺菌種ATCC 20888突變體係用PLR85、經由帶有酵 素預處理的電穿孔予以轉形（參見實施例8)，PLR85含有密 碼子最佳化的破囊壺菌種ATCC PTA-10212 序列辨modify. The flask was incubated on an shaker at 3 Torr and approximately 100 rpm for 4 h. The cells are monitored under a microscope to determine the extent of protoplasting, with the desired single cell. The cells were centrifuged at approximately 2500 xg for 5 min in a round bottom tube (i.e., l4inLFaiconTM tube, BD Biosciences, San Jose, CA). The suspension was removed and the cell lines were gently resuspended with 5 mL of ice-cold 10% glycerol. The cells were again centrifuged at approximately 2500 X g for 5 min in a round bottom tube. The suspension was removed and the cell lines were gently resuspended with 500 pL of ice-cold 10% glycerol using a wide-bore micropipette. The cell fraction of 9 〇 pL was placed in a pre-cooled electro-optical tube (Gene Pulser® cuvette _〇.1 cm gap or 〇.2 cm gap, Bio-Rad, Hercules, CA). One to five pg of DNA (at a volume of less than or equal to 10 pL) was added to the cuvette, gently mixed with a micropipette' and placed on ice for 5 min. Cell lines were electroporated at 200 〇 s (resistance), 25 pF (capacitance), and 250 V (for 0.1 cm gap) or 5 〇〇 v (〇. 2 cm gap). Add 0.5 mL of M50-20 medium to the cuvette immediately. The cells were then transferred to 4.5 mL of M50-20 medium in a 25 mL shake flask and incubated on a shaker at 2-3 ° C and approximately 100 rpm for 2-3 h. The cells were centrifuged at approximately 2500 X g for 5 m i η in a round bottom tube. The suspension was removed and the cell pellet was resuspended in 5 mL of Μ50-20 medium. The cells were plated on a suitable number (2 to 5) of M2B plates and incubated with appropriate selection and incubation at 3 °C. 139 201038734 The Schizochytrium strain ATCC 2〇888 mutant lacking functional orfA was also transformed with PREZ345 containing PFW, whereby the Keke 7 line was randomly combined with the mutant and restored PUFA production. Example 8 Thraustochytrium sp. ATCC Ding Ba-1〇212/^7 (SEQ ID NO: 68) was resynthesized (DNA2.0) and codon optimized for performance in Schizochytrium (Sequence ID: 120) 'and was selected to be expressed within a performance vector to generate pLR95. Codon optimization was generated using the Schizochytrium codon usage table in Figure 22. The expression vector contains approximately 2 kb of DNA from the flanking region of the native orfA gene locus of Schizochytrium sp. ATCC 20888. The Schizochytrium sp. ATCC 20888 mutant system from Example 7 lacking functional orfA uses a codon-optimized Thraustochytrium sp. ATCC ΡΤΑ-10212ΡΛ4/ (SEQ ID NO: 120) pLR95, via The electroporation of the enzyme pretreatment was transformed (see Example 7). According to the side of the anti-GiomycinTM marker in the mutant and the homologous region of the side of the Huko 7 gene in pLR95, double-exchange recombination occurs, whereby the codon-optimized cyst The genus ATCC ΡΤΑ-1〇212 ke/system was inserted into the natural orfA position. Recombination of the codon-optimized Thraustochytrium sp. ATCC PTA-10212 cadaver E4J (SEQ ID NO: 120) restored the production of PUFA in the absence of orfA of the Schizochytrium sp. ATCC 20888 mutant. Cell lines were grown and analyzed for FAMEs as described in Example 7. The naturally occurring Schizochytrium sp. ATCC 20888 strain containing the functional orfA gene produced a ratio of DHA and EPA of 25:1. 140.38734 codon-optimized species of Thraustochytrium sp. ATCC ΡΤΑ-10212, which replaces the non-activated orfA gene. Sequence identification number: 120) The recombinant strain produced a ratio of 5.4:1 DHA and EPA, further showing cracking The PUFA form of the bacterium can be altered by the nucleic acid molecules described herein. The recombinant strain had an EPA content of 4.4% FAME, a DPAn-3 content of 2.3%, a DPAn-6 content of 4.9%, and a DHA content of 24.0%. The Schizochytrium sp. ATCC 20888 mutant lacking functional orfA was also transformed with pLR95 containing it, thereby randomly binding to the mutant and restoring PUFA production. Example 9 The native orfB gene in Schizochytrium sp. ATCC 20888 was replaced by homologous recombination, followed by transformation with a vector via electroporation with enzyme pretreatment, containing the vector from orffl. The anti-GiomycinTM marker is surrounded by a sequence of lateral regions. A mutant strain lacking the functional orfB gene is produced. This mutant strain is auxotrophic and requires PUFA supplementation for growth. The Schizochytrium strain ATCC PTA-9695 corpse/^ (SEQ ID NO: 3) was cloned into the expression vector pDS 04 to generate pREZ3 31. The expression vector contained approximately 2 kb of DNA from the flanking region of the native orfB gene position of Schizochytrium sp. ATCC 20888. The Schizochytrium sp. ATCC 20888 mutant system lacking functional orfB was transformed with pREZ331 containing PFA2. Based on the random combination in this mutant, PUFA production was restored by Schizochytrium sp. ATCC PTA-9695 Coke 2 (serial identification number: 3). The cell lines were grown as described in Example 7 141 201038734 and analyzed for FAMEs. The natural Schizochytrium sp. ATCC 20888 strain containing the functional orfB gene produced dha and DPA n-6 in a ratio of 2.3:1. The recombinant strain containing the Schizochytrium sp. ATCC PTA_9695 (SEQ ID NO: 3) containing the uninactivated orfB gene produced a ratio of DHA and DPA n-6 of 3.5:1. The recombinant strain had a hydrazine content of 〇 8 〇 / FA FAME, a DPA η-3 content of 0.1% ' DPA n-6 content of 7.1%, and a DHA content of 25.1%. The Schizochytrium sp. ATCC 20888 mutant lacking functional orfB was also transformed with pREZ331 containing PFW, thereby inserting into the native orfB position and restoring PUFA production. Example 10 Thraustochytrium sp. ATCC PTA-10212 PFA2 (SEQ ID NO: 70) was resynthesized (DNA2.0) and codon optimized for expression in Schizochytrium (sequence identification number: 121) And being colonized into a performance vector to produce PLR85. Codon optimization was generated using the Schizochytrium codon usage table in Figure 22. The expression vector contains approximately 2 kb of DNA from the flanking region of the native orfB gene locus of Schizochytrium sp. ATCC 20888. Substitution of the orf gene was also investigated in Schizochytrium sp. ATCC 20888 with improved DHA productivity. The native orfB gene in the progeny strain is replaced by homologous recombination, followed by transformation with a vector via electroporation with enzyme pretreatment (see Example 7) containing the side from orffl The anti-GiomycinTM marker is surrounded by a sequence of regions. A mutant strain lacking the functional orfB gene was generated. The mutation 142 201038734 strain is auxotrophic and requires PUFA supplementation for growth. The mutant strain was transformed with pLR85 via electroporation with enzyme pretreatment (see Example 8) containing codon-optimized Thraustochytrium sp. ATCC PTA-10212 PE42 (SEQ ID NO: 121 ). According to the side of the anti-GiomycinTM marker in the mutant and the homologous region of the side of the PM2 gene in pLR85, double-exchange recombination occurs, whereby codon-optimized Thraustochytrium The ATCCPTA-l〇212 necropsy sequence number: 121) was inserted into the native orfB position of the mutant strain. Recombination with the codon-optimized Thraustochytrium strain ATCC ΡΤΑ-10212 Sequence Identification No.: 121) restored the production of PUFA in the absence of orfB progeny mutants. Cell lines were grown and analyzed for FAMEs as described in Example 7. The recombinant strain had an EPA content of 1.0% FAME, a DPAn-3 content of 0.3%, a DPAn-6 content of 7.0%, and a DHA content of 31.0%. In an experiment to be performed, the Schizochytrium sp. ATCC 20888 mutation system from Example 9 lacking functional orfB was transformed with PLR85 via electroporation with enzyme pretreatment (see Example 8), PLR85 Sequence analysis of Thraustochytrium sp. ATCC PTA-10212 containing codon optimization

s线編號.121)。根據在該突變體内的抗吉歐黴素tm標誌之 側邊以及在pLR85内的尸抑2基因的側邊之同源的區域，發 生雙交換重組，藉此密碼子最佳化的破囊壺菌種ATCC PTA-10212 (序列辨識編號：丨21)係插入至天然的orfB 位置中。具有密碼子最佳化的破囊壺菌種ATCC PTA_1〇2i2 (序列辨識編號：121)之重組作用恢復了在缺少〇rfB之 143 201038734 裂殖壺菌種ATCC 20888突變體内之PUFA的生產。 缺少功能性orffl之裂殖壺菌種ATCC 20888和後代菌株 突變體也用含有户柯2的pLR85予以轉形，藉此户柯2係隨機 與該突變體結合在一起以及恢復該等突變體中的各個之 PUFA的生產。 實施例11 一含有於裂殖壺菌内之抗巴龍黴素(paromomycin)標誌、 卡匣功能性的質體係發展出用於裂殖壺菌種ATCC 20888，其係藉由用原始來自細菌轉位子Τη5的新黴素磷酸 轉移酶Il(npt)之取代pMON50000/pTUBZEO 11 -2内的博歐 黴素/抗吉歐黴素TM基因（ble)編碼區域（美國專利7,001，772 B2)。於pMON50000中’ble抗性基因係由裂殖壺菌α-微管 蛋白啟動子所驅動以及接著SV40轉錄終止子。ρΜΟΝ50000 中的ble區域包含在ATG起始密碼子處的Ncol限制性位址以 及立即接在TGA停止訊息後之Pmll限制性位址。PCR係用 來擴增pCaMVnpt中存在的npt編碼區域(Shimizu等人，Plant J. 26(4):375 (2001))，藉此該產物包括在開始的ATG(粗體) 處之BspHI限制性位址（下面劃線的，引子CAX055)以及一 立即接在停止訊息（粗體-反向互補的）後的Pmll限制性位 址（下面劃線的，引子CAX056): CAX055(順向）： GTCATGATTGAACAAGATGGΑΤΤΠΓaγ (序列辨識 編號：66) CAX056(反向）： 144 201038734 C CACGTGTC AG A A G A A CTC GIC A AG A A (序列辨 識編號：67)。 PCR係用TaqMaster聚合酶套組(5Prime)來進行，產物係 被選殖至pCR4-TOPO(Invitrogen)之内，以及形成的質體係 被轉形至大腸桿菌TOPIO (Invitrogen)内。使用載體引子之 DNA序列分析係鑑定出含有所欲的805bp結構之多殖株（亦 即，該等序列係與加上經改造的限制性位址之來源模板的 该等序列匹配）。經修飾的npt編碼區域係藉由BSpHI加上 Pmll限制轉的消化作用予以單離，以及該經純化的dna片 段係用PMON50000載體片段予以連接，該載體片段係用 Ncol加上pmlI酵素予以消化而產生的。限制酶BspH^〇Nc〇I 留下相容的重疊末端’以及Pmll留下鈍的末端。形成的質 體’pTS-NPT’於完全相同的環境中含有npt新黴素/巴龍黴 素基因抗性基因，如同PMON50000中的原始ble基因之該 者。 使用裂殖壺菌的粒子轟擊法(particle bombardment)(美 國專利7,001,772 B2)來評估pTS-NPT中之新穎的抗巴龍黴 素卡II的功能。抗巴龍黴素(PAR)的挑選係於含有5 〇 pg/mL 硫酸巴龍黴素(Sigma)的瓊脂平盤上進行。抗巴龍黴素的裂 殖壺菌轉形體係以如同來自pM〇N50000的抗吉歐黴素^*之 該等相似的頻率被找到。”α·微管蛋白啟動子/叩t/SV4〇終止 子”卡匣可以用各種各樣的限制酶而自pTS-NPT解除供用 於其他發展的努力之隨後的用途。 實施例12 145 201038734 裂殖壺菌種ATCC 20888内的天然的orfC基因係藉由同 源性重組予以取代，接著用一載體予以轉形，該載體含有 被來自orfC的側邊區域的序列所圍繞之抗巴龍黴素標誌。 產生缺少功能性orfC基因的一突變菌株。該突變菌株為營 養缺陷型的且需要用於生長之PUFA補充。 裂殖壺菌種ATCC PTA-9695 序列辨識編號：5) 係被選殖至表現載體pREZ22之内以產生pREZ324。該表現 載體含有來自裂殖壺菌種ATCC 20888之天然的orfC基因位 置的側邊區域之大概2 kb的DNA。 缺少功能性orfC的裂殖壺菌種ATCC 20888突變體係用 含有裂殖壺菌種ATCC PTA-9695户烈3的pREZ324予以轉 形。根據在該突變體内的抗巴龍黴素標誌的側邊以及在 pREZ324内的裂殖壺菌種ATCC PTA-9695 基因的側邊 之同源的區域，發生雙交換重組，藉此裂殖壺菌種ATCC PTA-9695 PFA3被插入至天然的orfC位置中。裂殖壺菌種 ATCC ΡΤΑ-9695 序列辨識編號：5)之同源性重組作用 恢復了在缺少orfC之裂殖壺菌種ATCC 20888突變體内之 PUFA的生產。細胞係如實施例7中說明的方式生長且分析 FAMEs。含有功能性orfC基因之天然的裂殖壺菌種ATCC 20888菌株係生產2.3 : 1的比率之DHA和DPA n-6。含有代 替不活化的orfC基因之裂殖壺菌種ATCC PTA-9695 (序列辨識編號：5)的重組型菌株生產14:9的比率之DHA和 DPA n-6，進一步展現出裂殖壺菌的PUFA形態可以藉由本 文中說明的核酸分子予以改變。該重組型菌株的EPA含量 146 201038734 為 1 _2°/〇 的 FAME，DPA n-3 含量為 〇 2%，dpa n-6含量為 2.9%，以及DHA含量為43.4%。 缺少功能性orfC之裂殖壺菌種ATCC 20888突變體也用 含有PFA3的pREZ324予以轉形，藉此PFA3係隨機結合於該 突變體内以及恢復PUFA的生產。該重組型菌株的epa含量 為 1.2°/。的 FAME，DPA n-3含量為 0.2%，DPA n-6含量為 2.5%，以及DHA含量為39.1%。 q 於實施例10中討論的後代菌株内之天然的orfC基因係 藉由同源性重組予以取代，接著用一載體予以轉形，該載 體含有被來自orfC的側邊區域的序列所圍繞之抗巴龍黴素 _ 標誌。產生缺少功能性orfC基因的一突變菌株。該突變菌 株為營養缺陷型的且需要用於生長之PUFA補充。缺少功能 ' 性orfC的突變體係用pREZ324予以轉形◊發生雙交換重組， 藉此裂殖壺菌種ATCC PTA-9695户抑3被插入至該突變菌 株之天然的orfC位置中。裂殖壺菌種ATCC PTA-9695 q (序列辨識編號：5)之同源性重組作用恢復了該缺少orfc之 後代菌株突變體内的PUFA的生產。細胞係如實施例7中說 明的方式生長且分析FAMEs。該重組型菌株的EPA含量為 1.2%的FAME，DPAn-3含量為 0.3%，DPAn-6含量為2.8%， 以及DHA含量為43.1%。 該缺少功能性〇 r ffl之後代菌株突變體也用含有户抱3之 PREZ324予以轉形，藉此係隨機與該突變體結合在一 起以及恢復PUFA的生產。 實施例13 147 201038734 破囊壺菌種ATCC PTA-10212户柯3 (序列辨識編號：72) 係被再合成(DNA2.0)且被密碼子最佳化為了於裂殖壺菌内 表現(序列辨識編號：122)，以及被選殖至表現載體pREZ22 之内以產生PREZ337。密碼子最佳化係使用第22圖中的裂 殖壺菌密碼子使用表來產生。該表現載體含有來自裂殖壺 菌種ATCC 20888的天然的orfC基因位置之側邊區域的大概 2 kb 的 DNA。 來自實施例12之缺少功能性orfC的後代菌株突變體係 用含有密碼子最佳化的破囊壺菌種ATCC PTA-10212尸 (序列辨識編號：122)之pREZ337、經由帶有酵素預處理的 電穿孔予以轉形（參見實施例8)。根據在該突變體内的抗吉 歐黴素TM標誌之側邊以及在PREZ337内的基因的側邊 之同源的區域，發生雙交換重組，藉此密碼子最佳化的破 囊壺菌種ATCCPTA-l〇2l2尸柯3(序列辨識編號：122)係插 入至天然的orfC位置中。具有密碼子最佳化的破囊壺菌種 ATCCPTA-10212PFd3(序列辨識編號：122)之重組作用恢 復了該缺少orfC之後代菌株突變體内的PUFA的生產。細胞 係如實施例7中說明的方式生長且分析F A Μ E s。該重組型菌 株的ΕΡΑ含量為1.3%的FAME，DPA η-3含量為0.4%，DPA η-6含量為2.7%，以及DHA含量為50.2%。 於要執行的一實驗中，來自實施例12之缺少功能性 orfC的裂殖壺菌種ATCC 20888突變體係用含有密碼子最佳 化的破囊壺菌種ATCC ΡΤΑ-1〇212尸柯3 (序列辨識編號： 122)之pREZ337、經由帶有酵素預處理的電穿孔予以轉形 148 201038734 (參見實施例8)。根據在該突變體内的抗吉歐黴素TM標誌之 側邊以及在PREZ337内的户Ε43基因的側邊之同源的區域， 發生雙交換重組，藉此密碼子最佳化的破囊壺菌種ATCC PTA-10212户柯3 (序列辨識編號：122)係插入至天然的orfC 位置中。具有密碼子最佳化的破囊壺菌種ATCC PTA-10212 序列辨識編號：122)之重組作用恢復了該缺少orfc之 裂殖壺菌種ATCC 20888突變體内的PUFA的生產。 缺少功能性orfC之裂殖壺菌種ATCC 20888以及後代菌 株突變體也用含有尸柯3的pREZ337予以轉形，藉此係 隨機與該突變體結合在一起且恢復該等突變體的各個中的 PUFA的生產。 實施例14 於裂殖壺菌種ATCC 20888内的orfA、orffl，和orfC基 因之任何2者或全部3者係藉由同源性重組予以取代，接著 用載體予以轉形，該載體含有被來自orf的側邊區域的序列 所圍繞之抗吉歐黴素⑽標誌或抗巴龍黴素標誌。缺少功能 性基因orfA、orfB ’和orfC的任何2者或全部3者之突變菌株 被產生。該等突變菌株為營養缺陷型的且需要用於生長之 PUFA補充。 缺少功能性orf基因之裂殖壺菌種ATCC 20888突變體 係用含有對應的PFA基因（序列辨識編號：1、3、5、120、 121 ’或是122的一或更多者)之一或更多個的表現載體予以 轉形。根據在該突變體内的抗吉歐黴素⑽標誌或抗巴龍黴 素標誌之側邊以及在各別的表現載體中的戶抑基因的側邊 149 201038734 之同源的區域，可發生雙交換重組，藉此户柯基因係插入至 天然的orf位置中。此等的表現載體的隨機結合亦能發生， 伴隨轉形體的挑選，僅根據PUFA的恢復。PFA基因之同源 性重組恢復了突變體中之PUFA的生產，藉此恢復天然的 PUFA形態或是根據被插入至突變體之内的PFA基因的組合 來改變天然的PUFA形態。 於一執行的實驗中，來自實施例12之裂殖壺菌種ATCC 20888菌株(缺少一功能性orfc基因以及含有隨機結合的裂 殖壺菌種八丁(^?丁八-9695户柯3(序列辨識編號：5))被用來 取代orfA基因和〇rfB基因。菌株中之天然的orfA基因和orfB 基因係藉由同源性重組予以取代，接著用一載體予以轉 形’該載體含有被來自orfA與orfB的側邊區域的序列所圍繞 之抗吉歐黴素TM標誌。一菌株係被產生，其缺少功能性 orfA、orffi，和orfC，以及含有隨機結合的裂殖壺菌種ATCC PTA-9695 該菌株係用含有密碼子最佳化的裂殖壺菌 種ATCC PTA-9695户柯7 (序列辨識編號：1)的pREZ345以及 含有密碼子最佳化的裂殖壺菌種ATCC PTA-9695 (序 列辨識編號：3)的pREZ331予以轉形，藉此發生尸與PE42 的隨機結合。形成的重組型菌株缺少官能性orfA、orfB，和 orfC以及含有隨機結合的裂殖壺菌種ATCC PTA-9695 、尸柯2，和戶E43。細胞係如實施例7中說明的方式生 長且分析FAMEs。該重組型菌株的EPA含量為6.6%的 FAME，DPA n-3含量為0.8%，DPA n-6含量為 1.6%，以及 DHA含量為20.9%。 150 201038734 於另一執行的實驗中，來自實施例12的後代菌株(缺少 一功能性orfC基因以及含有插入至天然的orfC位置中之裂 殖壺菌種ATCC PTA-9695 (序列辨識編號：5))被用來 取代orfA基因和orfB基因。菌株中之天然的orfA基因和 基因係藉由同源性重組予以取代，接著用一載體予以轉 形’該載體含有被來自orfA與orfB的側邊區域的序列所圍繞 之抗巴龍黴素標誌。一菌株係被產生，其缺少功能性orfA、 orfB ’和orfC ’以及含有插入至天然的orfC位置中的裂殖壺 菌種ATCC PTA-9695 PFJ3。該菌株係用含有密碼子最佳化 的裂殖壺菌種ATCC PTA-9695 序列辨識編號：丨）之 pREZ345以及含有密碼子最佳化的裂殖壺菌種ATCC PTA-9695尸柯2 (序列辨識編號：3)之pREZ331予以轉形。 發生雙交換重組，藉此裂殖壺菌種ATCC PTA-9695 PF以係 插入至菌株的天然的orfA位置中以及裂殖壺菌種ATCC PTA-9695 PE42係插入至菌株的天然的orfB位置中。形成的 重組型菌株缺少官能性orfA、orfB，和orfC，以及含有被插 入至各別的orfA、orfB，和orfC位置之内的裂殖壺菌種ATCC PTA-9695 PE4/、户柯2，以及。細胞係如實施例7中說 明的方式生長且分析FAMES。該重組型菌株的EPA含量為 7.3%的FAME，DPAn-3含量為0.4%，DPAn-6含量為 1.5%， 以及DHA含量為23.9%。 於另一執行的實驗中，來自實施例12的後代菌株(缺少 一功能性orfC基因以及含有隨機結合的裂殖壺菌種ATCC PTA-9695尸E43(序列辨識編號：5))被用來取代orfA基因和 151 201038734 orffi基因。菌株中之天然的orfA基因和orfB基因係藉由同源 性重組予以取代’接著用一載體予以轉形，該載體含有被 來自orfA與orfB的側邊區域的序列所圍繞之抗吉歐黴素tm 標遠'。一函株係被產生’其缺少功能性orfA、orfB，和orfC， 以及含有隨機結合的裂殖壺菌種ATCC PTA-9695 。該 菌株係用含有密碼子最佳化的裂殖壺菌種ATCC PTA-9695 (序列辨識編號：1)之PREZ345以及含有密碼子最佳化 的裂殖壺菌種ATCC PTA-9695 (序列辨識編號：3)之 pREZ331予以轉形，藉此發生p/⑷與的隨機結合。形 成的重組型菌株缺少官能性orfA、orfB，和orfC以及含有隨 機結合的裂殖壺菌種ATCC PTA-9695，和 PE43。細胞係如實施例7中說明的方式生長且分析FAMEs。 該重組型菌株的EPA含量為6.2%的FAME，DPA n-3含量為 1.3%，DPAn-6含量為 0.9%，以及DHA含量為 16.6%。 於另一執行的實驗中，來自實施例13的後代菌株(缺少 一功能性orfC基因以及含有插入至天然的orfC位置中之裂 殖壺菌種ATCC PTA-10212户柯3 (序列辨識編號：122))被用 來取代orfA基因和orfB基因。菌株中之天然的orfA基因和 orfB基因係藉由同源性重組予以取代，接著用一載體予以 轉形，該載體含有被來自orfA與orfB的側邊區域的序列所圍 繞之抗巴龍黴素標誌。一菌株係被產生，其缺少功能性 orfA、orfB，和orfC，以及含有插入至天然的orfC位置中的 裂殖壺菌種ATCC PTA-10212 。該菌株係用含有密碼 子最佳化的裂殖壺菌種ATCC PTA-10212 (序列辨識 152 201038734 編號：120)之pLR95以及含有密碼子最佳化的裂殖壺菌種 ATCC PTA-10212 (序列辨識編號：121)之pLR85予以 轉形。發生雙交換重組，藉此裂殖壺菌種ATCCPTA-10212 PiM Η系插入至菌株的天然的orfA位置中以及裂殖壺菌種 ATCC PTA-10212 PE42係插入至菌株的天然的〇rfB位置 中。形成的重組型菌株缺少官能性orfA、orfB，和orfC，以 及含有被插入至各別的orfA、orfB，和orfC位置之内的裂殖 壺菌種ATCC PTA-10212尸FA、，以及尸柯3。細胞係 如實施例7中說明的方式生長且分析f a Μ E s。該重組型菌株 的ΕΡΑ含量為5.2%的FAME，DPAn-3含量為0.6%，DPAn-6 含量為2.1%，以及DHA含量為47.1%。 於另一執行的實驗中，來自實施例13的後代菌株(缺少 一功能性orfC基因以及含有隨機結合的裂殖壺菌種ATCC PTA-10212PFJJ(序列辨識編號：丨22))被用來取代orfA基因 和orfB基因。菌株中之天然的orfA基因和orfB基因係藉由同 源性重組予以取代’接著用一載體予以轉形，該載體含有 被來自orfA與orfB的側邊區域的序列所圍繞之抗吉歐黴素 TM標誌、。一菌株係被產生，其缺少功能性orfA、〇rfB，和 orfC ’以及含有隨機結合的裂殖壺菌種ATCC PTA-10212 。該菌株係用含有密碼子最佳化的裂殖壺菌種ATCC PTA-10212 PFW (序列辨識編號：i2〇)之pLR95以及含有密 碼子最佳化的裂殖壺菌種ATCC PTA-10212 序列辨 識編號：121)之pLR85予以轉形，藉此發生/與户柯2的 隨機結合。形成的重組型菌株缺少官能性orfA、orfB，和orfc 153 201038734 以及含有隨機結合的裂殖壺菌種ATCC PTA-10212尸抑7、 PFX2 ’以及PF/13。細胞係如實施例7中說明的方式生長且 分析FAMEs。該重組型菌株的ΕΡΑ含量為1.8%的FAME， DPAn-3含量為1.8%，DPAn-6含量為2.3%，以及DHA含量 為 34.1%。 實施例15 來自裂殖壺菌種ATCC 20888的orfA、orffl，和orfC基 因係被選殖至一系列的Duet載體(Novagen)之内。Duet表現 載體為一組相容的質體，其中多標的基因係被選殖以及由 大腸桿菌内的T7誘導性啟動子予以共表現。Duet質體 pREZ91 含有裂殖壺菌種 ATCC 20888 orfA 於 pETDuet-1 内；duet質體pREZ96含有裂殖壺菌種ATCC 20888 orffl於 pCDFDuet-1内；以及duet質體pREZlOl含有裂殖壺菌種 ATCC 20888 orfC於pCOLADuet-1 内。Duet質體pREZ91、 pREZ96,以及pREZlOl，與含有需要的輔助基因Hetl質體 pJK737 (於美國專利案第7,217,856號之中說明，以其之整 體併入本文中作為參考資料）一起’係被轉形至大腸桿菌菌 株BLR (DE3)内，其含有一誘導性T7 RNA聚合酶基因。一 旦細胞生長以及添加IPTG ’依據製造者之指示(Novagen)， DHA和DPAn-6係被生產。簡言之’當細胞達到於600 nm之 大約0.5的光密度時，添加1 mM IPTG用於誘導。細胞於 Luria肉湯内在3(TC下生長12小時以及收穫。該等脂肪酸係 使用標準的技術轉變成甲酯。脂肪酸形態係使用附有火焰 離子偵測器之氣相層析（GC_FID)決定為脂肪酸甲酯 154 201038734 (FAME)。 裂殖壺菌種ATCC PTA-9695尸FW (序列辨識編號：i) 基因係被選殖至該表現載體pETDuet-1之内，產生 PREZ346。Duet質體pREZ346 (含有裂殖壺菌種ATCC PTA-9695 戶E4/)、pREZ96 (含有orffi)，以及pREZlOl(含有 orfC)係與pJK737 (含有Hetl)—起被轉形至大腸桿菌菌株 801(0£3)内。裂殖壺菌種八丁(^?丁八-9695 />/^基因係與 裂殖壺菌種ATCC 20888 orfB和orfC基因共表現。裂瘦壺菌 種ATCC PTA-9695 ，組合以裂殖壺菌種ATCC 20888 orffl與orfC之表現’支持誘導條件下之大腸桿菌内的dha 的生產。該經轉形的大腸桿菌之DHA含量為2.8%的 FAME，DPA n-6含量為 1.1%，DPA n-3含量為0.6%，以及 EPA含量為3.7%。 實施例16 密碼子最佳化的破囊壺菌種ATCC PTA-10212户 (序列辨識編號：120)基因係被選殖至該表現載體 pETDuet-Ι之内’產生pLRIOO。Duet質體pLRl00 (含有密碼 子最佳化的破囊壺菌種ATCC PTA-10212 PE4/)、PREZ96 (含有裂殖壺菌種ATCC 20888 orfB)，以及pREZlOl (含有裂 殖壺菌種ATCC 20888 orfC)係與pJK737 (含有Hetl)—起被 轉形至大腸桿菌菌株BLR(DE3)内。參見實施例15。破囊壺 菌種ATCC PTA-10212户柯7基因係與裂殖壺菌種ATcc 2〇888 orfB和orfC基因共表現。破囊壺菌種ATCC PTA-10212 PiM/ ’組合以裂殖壺菌種ATCC 20888 orffl與 155 201038734 orfC之表現，支持誘導條件下之大腸桿菌内的^^八與EpA 的生產。 實施例17 裂殖壺菌種ATCC PTA-9695尸(序列辨識編號：5) 基因係被選殖至該表現載體pCOLADueH之内，產生 pREZ326。Duet質體pREZ326 (含有裂殖壺菌種ATCC PTA-9695 PFW)、pREZ91 (含有裂殖壺菌種ATCc 20888 orfA) ’以及pREZ96 (含有裂殖壺菌種ATCC 2〇888 〇rfB)係 與pJK737 (含有Hetl)—起被轉形至大腸桿菌菌株BLR(DE3) 内。參見實施例15。裂殖壺菌種ATCC ΡΤΑ-9695 ，組 合以裂殖壺菌種ATCC 20888 orfA和orfB的表現，支持誘導 條件下之大腸桿菌内的DHA的生產。細胞係如實施例15中 說明的方式生長且分析FAMEs。該經轉形的大腸桿菌之 DHA含量為〇·3%的FAME。 實施例18 密碼子最佳化的破囊壺菌種ATCC PTA-10212 (序列辨識編號：122)基因係被選殖至該表現載體 pCOLADuet-Ι之内，產生pREZ348°Duet質體pREZ34(含有 密碼子最佳化的破囊壺菌種ATCC PTA-10212 PE43)、 pREZ91 (含有裂殖壺 g 種ATCC 20888 orfA)，以及pREZ96 (含有裂殖壺菌種ATCC 20888 orfB)係與PJK737 (含有Hetl) 一起被轉形至大腸桿菌菌株BLR (DE3)内。參見實施例15。 破囊壺菌種ATCC PTA-l〇2l2 ，組合以裂殖壺菌種 ATCC 20888 orfA和orfB的表現，支持誘導條件下之大腸桿 156 201038734 菌内的DHA的生產。細胞係如實施例15中說明的方式生長 且分析FAMEs。該經轉形的大腸桿菌之DHA含量為2 9%的 FAME，以及該DPAn-6含量為0_4〇/〇。 實施例19 裂殖壺菌種ATCC PTA-9695 PE42 (序列辨識編號：3) 基因係被選殖至該表現載體pCDFDuet-Ι之内，產生s line number.121). According to the side of the anti-getomycin tm marker in the mutant and the homologous region of the side of the cadaver 2 gene in pLR85, double-crossover recombination occurs, whereby codon-optimized cysts are generated. The genus ATCC PTA-10212 (SEQ ID NO: 丨 21) was inserted into the native orfB position. The recombination of the codon-optimized Thraustochytrium sp. ATCC PTA_1〇2i2 (SEQ ID NO: 121) restored the production of PUFA in the absence of 〇rfB 143 201038734 Schizochytrium ATCC 20888 mutant. Schizochytrium strain ATCC 20888 lacking functional orffl and mutants of progeny strains were also transformed with pLR85 containing Hu Ke 2, whereby the Keke 2 line was randomly associated with the mutant and restored to the mutants. The production of each PUFA. Example 11 A paromomycin-containing and calyx functional system contained in Schizochytrium was developed for Schizochytrium sp. ATCC 20888, which was transferred from the original bacteria. The neomycin phosphotransferase Il (npt) of the position Τn5 is substituted for the Boomycin/anti-GiomycinTM gene (ble) coding region in pMON50000/pTUBZEO 11 -2 (U.S. Patent 7,001,772 B2). The 'ble resistance gene line in pMON50000 is driven by the Schizochytrium alpha-tubulin promoter followed by the SV40 transcriptional terminator. The ble region in ρΜΟΝ50000 contains the Ncol-restricted address at the ATG start codon and the Pmll-restricted address immediately after the TGA stop message. The PCR system was used to amplify the npt coding region present in pCaMVnpt (Shimizu et al., Plant J. 26(4): 375 (2001)) whereby the product included BspHI restriction at the beginning of ATG (bold) The address (underlined, the primer CAX055) and a Pmll-restricted address immediately after the stop message (bold-reverse complement) (underlined, primer CAX056): CAX055 (forward): GTCATGATTGAACAAGATGGΑΤΤΠΓaγ (SEQ ID NO: 66) CAX056 (reverse): 144 201038734 C CACGTGTC AG AAGAA CTC GIC A AG AA (Sequence Identification Number: 67). The PCR system was carried out using the TaqMaster polymerase kit (5 Prime), and the product was cloned into pCR4-TOPO (Invitrogen), and the resulting system was transformed into E. coli TOPIO (Invitrogen). DNA sequence analysis using vector primers identified multiple colonies containing the desired 805 bp structure (i.e., the sequences were matched to the sequences of the source template with the modified restriction site). The modified npt coding region was isolated by digestion with BpHI plus Pmll restriction, and the purified DNA fragment was ligated with the PMON50000 vector fragment, which was digested with Ncol plus pmlI enzyme. produced. The restriction enzyme BspH^〇Nc〇I leaves a compatible overlapping end' and Pmll leaves a blunt end. The resulting plasmid 'pTS-NPT' contains the npt neomycin/balonmycin gene resistance gene in exactly the same environment as the original ble gene in PMON50000. The function of the novel anti-Baronin card II in pTS-NPT was evaluated using the particle bombardment of Schizochytrium (U.S. Patent 7,001,772 B2). Selection of anti-paromomycin (PAR) was performed on an agar plate containing 5 〇 pg/mL of paromomycin sulfate (Sigma). The anti-paronimycin-derived Schizochytrium transformation system was found at such similar frequencies as anti-Giomycin® from pM〇N50000. The "α·tubulin promoter/叩t/SV4〇 terminator” can be depleted from pTS-NPT by a variety of restriction enzymes for subsequent use in other development efforts. Example 12 145 201038734 The native orfC gene within Schizochytrium sp. ATCC 20888 was replaced by homologous recombination followed by transformation with a vector containing sequences sequenced from the flanking regions of orfC. Anti-paromomycin logo. A mutant strain lacking the functional orfC gene is produced. The mutant strain is nutritionally deficient and requires PUFA supplementation for growth. Schizochytrium strain ATCC PTA-9695 Sequence ID: 5) was cloned into the expression vector pREZ22 to generate pREZ324. The expression vector contains approximately 2 kb of DNA from the flanking region of the natural orfC gene position of the Schizochytrium sp. ATCC 20888. The Schizochytrium sp. ATCC 20888 mutant system lacking functional orfC was transformed with pREZ324 containing Schizochytrium sp. ATCC PTA-9695 Tore 3 . According to the side of the anti-paromomycin marker in the mutant and the homologous region of the side of the Schizochytrium sp. ATCC PTA-9695 gene in pREZ324, double-crossover recombination occurs, whereby the fission pot The strain ATCC PTA-9695 PFA3 was inserted into the native orfC position. Schizochytrium strain ATCC ΡΤΑ-9695 Sequence ID: 5) Homology recombination The production of PUFA in the absence of orfC of the Schizochytrium sp. ATCC 20888 mutant was restored. Cell lines were grown and analyzed for FAMEs as described in Example 7. The natural Schizochytrium sp. ATCC 20888 strain containing the functional orfC gene produced DHA and DPA n-6 in a ratio of 2.3:1. A recombinant strain containing the Schizochytrium sp. ATCC PTA-9695 (SEQ ID NO: 5) in place of the inactivated orfC gene produces a 14:9 ratio of DHA and DPA n-6, further exhibiting Schizochytrium The PUFA morphology can be altered by the nucleic acid molecules described herein. The recombinant strain had an EPA content of 146 201038734 of 1 _2 ° / 〇 FAME, DPA n-3 content of 〇 2%, dpa n-6 content of 2.9%, and DHA content of 43.4%. The Schizochytrium sp. ATCC 20888 mutant lacking functional orfC was also transformed with pREZ324 containing PFA3, whereby the PFA3 line was randomly bound to the mutant and the production of PUFA was resumed. The recombinant strain had an epa content of 1.2 ° /. The FAME had a DPA n-3 content of 0.2%, a DPA n-6 content of 2.5%, and a DHA content of 39.1%. q The native orfC gene in the progeny strain discussed in Example 10 is replaced by homologous recombination followed by transformation with a vector containing an antibody surrounded by sequences from the flanking regions of orfC. Paromomycin _ logo. A mutant strain lacking the functional orfC gene is produced. The mutant strain is auxotrophic and requires PUFA supplementation for growth. Lack of function 'Sexual orfC mutant system was transformed with pREZ324 to generate double-crossover recombination, whereby Schizochytrium sp. ATCC PTA-9695, 3, was inserted into the natural orfC position of the mutant strain. The homologous recombination of the Schizochytrium sp. ATCC PTA-9695 q (SEQ ID NO: 5) restored the production of PUFA in the mutant of the progeny strain lacking orfc. Cell lines were grown and analyzed for FAMEs as described in Example 7. The recombinant strain had an EPA content of 1.2% FAME, a DPAn-3 content of 0.3%, a DPAn-6 content of 2.8%, and a DHA content of 43.1%. The lack of functional 〇 r ffl progeny strain mutants were also transformed with PREZ324 containing the cultivar 3, thereby randomly combining with the mutant and restoring PUFA production. Example 13 147 201038734 Thraustochytrium sp. ATCC PTA-10212 Hu Ke 3 (SEQ ID NO: 72) was resynthesized (DNA2.0) and codon optimized for expression in Schizochytrium (sequence) Identification number: 122), and was selected to be within the performance vector pREZ22 to generate PREZ337. Codon optimization was generated using the Schizochytrium codon usage table in Figure 22. The expression vector contains approximately 2 kb of DNA from the flanking region of the native orfC gene locus of the Schizochytrium sp. ATCC 20888. The progeny strain mutant system from Example 12 lacking functional orfC was subjected to codon-prepared Thraustochytrium sp. ATCC PTA-10212 (SEQ ID NO: 122) pREZ337, via enzyme pretreated The perforations were transformed (see Example 8). A double-recombinant recombination based on the side of the anti-geomycinTM marker in the mutant and the homologous region of the gene in the PREZ337, whereby the codon-optimized species ATCCPTA-l〇2l2 nectar 3 (sequence identification number: 122) was inserted into the native orfC position. Recombination of the codon-optimized Thraustochytrium strain ATCCPTA-10212PFd3 (SEQ ID NO: 122) restored the production of PUFAs in the absence of orfC progeny strain mutants. The cell lines were grown and analyzed for F A Μ E s as described in Example 7. The recombinant strain had a terpene content of 1.3% FAME, a DPA η-3 content of 0.4%, a DPA η-6 content of 2.7%, and a DHA content of 50.2%. In an experiment to be performed, the Schizochytrium sp. ATCC 20888 mutant system lacking functional orfC from Example 12 was codon-preserved with a codon-optimized species: ATCC ΡΤΑ-1〇212 necco 3 ( Sequence identification number: 122) pREZ337, transformed via electroporation with enzyme pretreatment 148 201038734 (see Example 8). A double-exchange recombination occurs according to the side of the anti-GiomycinTM marker in the mutant and the homologous region of the side of the Toza 43 gene in PREZ337, whereby the codon-optimized The strain ATCC PTA-10212 Hu Ke 3 (SEQ ID NO: 122) was inserted into the native orfC position. Recombination with the codon-optimized Thraustochytrium sp. ATCC PTA-10212 Sequence ID: 122) restored the production of PUFAs in the ATCC 20888 mutant of the Schizochytrium strain lacking orfc. Schizochytrium strain ATCC 20888 lacking functional orfC and mutant strains of progeny strains were also transformed with pREZ337 containing nectar 3, whereby the mutant was randomly combined with the mutant and recovered in each of the mutants. Production of PUFA. Example 14 Any two or all of the orfA, orffl, and orfC genes in Schizochytrium sp. ATCC 20888 were replaced by homologous recombination, followed by transformation with a vector containing the vector from The sequence of the side regions of orf is surrounded by an anti-Giomycin (10) marker or an anti-paromomycin marker. A mutant strain lacking any two or all of the functional genes orfA, orfB' and orfC was produced. These mutant strains are auxotrophic and require PUFA supplementation for growth. The Schizochytrium strain ATCC 20888 mutant system lacking the functional orf gene uses one or more of the corresponding PFA genes (sequence identification number: 1, 3, 5, 120, 121 ' or 122) or more A plurality of performance carriers are transformed. A double may occur depending on the side of the anti-Giomycin (10) marker or the anti-paromomycin marker in the mutant and the homologous region of the side 149 201038734 of the house suppressor gene in the respective expression vector. Exchange recombination, whereby the genomic gene is inserted into the native orf position. Random binding of such performance vectors can also occur, with the selection of transformants, based solely on the recovery of PUFAs. Homologous recombination of the PFA gene restores the production of PUFAs in the mutant, thereby restoring the native PUFA morphology or altering the native PUFA morphology based on the combination of PFA genes inserted into the mutant. In an experiment performed, the Schizochytrium sp. ATCC 20888 strain from Example 12 (lack of a functional orfc gene and a sclerotium sclerotium containing a random binding) (^? D-8-9695 ke 3 (sequence) Identification number: 5)) is used to replace the orfA gene and the 〇rfB gene. The natural orfA gene and orfB gene in the strain are replaced by homologous recombination, followed by transformation with a vector containing the vector from An anti-GiomycinTM marker surrounding the sequence of the side regions of orfA and orfB. A strain is produced that lacks functional orfA, orffi, and orfC, and a Schwann sp. ATCC PTA-containing random binding. 9695 This strain uses pREZ345 containing the codon-optimized Schizochytrium strain ATCC PTA-9695, Keke 7 (SEQ ID NO: 1) and the codon-optimized Schizochytrium strain ATCC PTA-9695 (SEQ ID NO: 3) pREZ331 was transformed, whereby random binding of cadaver to PE42 occurred. The resulting recombinant strain lacked functional orfA, orfB, and orfC and the Schwann sp. ATCC PTA- containing random binding. 9695, corpse 2, and household E43. Fine The FAMEs were grown and analyzed as described in Example 7. The recombinant strain had an EPA content of 6.6% FAME, a DPA n-3 content of 0.8%, a DPA n-6 content of 1.6%, and a DHA content of 20.9. 150 201038734 In another performed experiment, the progeny strain from Example 12 (lack of a functional orfC gene and the Schizochytrium species ATCC PTA-9695 containing the position inserted into the native orfC position (SEQ ID NO: 5)) is used to replace the orfA gene and the orfB gene. The natural orfA gene and gene in the strain are replaced by homologous recombination, followed by transformation with a vector containing the side from orfA and orfB The anti-paromomycin marker surrounding the sequence of the border region. A strain is produced which lacks functional orfA, orfB 'and orfC ' and the Schizochytrium strain ATCC PTA-9695 containing the position into the native orfC position. PFJ3. This strain uses the codon-optimized Schizochytrium strain ATCC PTA-9695 sequence identification number: 丨) of pREZ345 and the codon-optimized Schizochytrium strain ATCC PTA-9695 necco 2 (sequence identification number: 3) p REZ331 is transformed. Double-crossover recombination occurs whereby Schizochytrium sp. ATCC PTA-9695 PF is inserted into the native orfA position of the strain and the Schizochytrium strain ATCC PTA-9695 PE42 is inserted into the native orfB position of the strain. The resulting recombinant strain lacks functional orfA, orfB, and orfC, as well as Schizochytrium species ATCC PTA-9695 PE4/, Hu Ke 2, which are inserted into the respective orfA, orfB, and orfC positions, and . The cell lines were grown and analyzed for FAMES as described in Example 7. The recombinant strain had an EPA content of 7.3% FAME, a DPAn-3 content of 0.4%, a DPAn-6 content of 1.5%, and a DHA content of 23.9%. In another performed experiment, the progeny strain from Example 12 (lack of a functional orfC gene and the mitochondrial strain ATCC PTA-9695 corpse E43 (SEQ ID NO: 5) containing the random binding) was used instead. OrfA gene and 151 201038734 orffi gene. The native orfA gene and the orfB gene in the strain are replaced by homologous recombination' followed by transformation with a vector containing anti-Gytomycin surrounded by sequences from the flanking regions of orfA and orfB. Tm standard far. A functional line was produced 'which lacks functional orfA, orfB, and orfC, and contains a randomly binding Schizochytrium strain ATCC PTA-9695. This strain uses the codon-optimized Schizochytrium strain ATCC PTA-9695 (SEQ ID NO: 1) PREZ345 and the codon-optimized Schizochytrium strain ATCC PTA-9695 (sequence identification number) :3) The pREZ331 is transformed, whereby a random combination of p/(4) and is generated. The resulting recombinant strain lacks functional orfA, orfB, and orfC as well as the Schwanchys sp. ATCC PTA-9695, and PE43 containing the random binding. Cell lines were grown and analyzed for FAMEs as described in Example 7. The recombinant strain had an EPA content of 6.2% FAME, a DPA n-3 content of 1.3%, a DPAn-6 content of 0.9%, and a DHA content of 16.6%. In another experiment performed, the progeny strain from Example 13 (lack of a functional orfC gene and the Schizochytrium species ATCC PTA-10212 containing the inserted into the native orfC position) (SEQ ID NO: 122 )) was used to replace the orfA gene and the orfB gene. The native orfA gene and the orfB gene in the strain are replaced by homologous recombination, followed by transformation with a vector containing anti-paromomycin surrounded by sequences from the flanking regions of orfA and orfB. Sign. A strain is produced which lacks the functional orfA, orfB, and orfC, as well as the Schizochytrium sp. ATCC PTA-10212 which is inserted into the native orfC position. This strain uses pLR95 containing the codon-optimized Schizochytrium strain ATCC PTA-10212 (SEQ ID NO: 152 201038734 number: 120) and the codon-optimized Schizochytrium strain ATCC PTA-10212 (sequence Identification number: 121) pLR85 is transformed. A double-crossover recombination occurs whereby the Schizochytrium sp. ATCCPTA-10212 PiM tether is inserted into the native orfA position of the strain and the Schizochytrium strain ATCC PTA-10212 PE42 is inserted into the native 〇rfB position of the strain. The resulting recombinant strain lacks functional orfA, orfB, and orfC, and contains Schizochytrium sp. ATCC PTA-10212 cadaver FA, which is inserted into the respective orfA, orfB, and orfC positions, and corpse 3 . Cell lines were grown and analyzed for f a Μ E s as described in Example 7. The recombinant strain had a hydrazine content of 5.2% of FAME, a DPAn-3 content of 0.6%, a DPAn-6 content of 2.1%, and a DHA content of 47.1%. In another experiment performed, the progeny strain from Example 13 (lack of a functional orfC gene and a Schwanchy sp. ATCC PTA-10212PFJJ (sequence identification number: 丨22) containing random binding) was used to replace orfA. Gene and orfB genes. The native orfA gene and the orfB gene in the strain are replaced by homologous recombination' followed by transformation with a vector containing anti-Gytomycin surrounded by sequences from the flanking regions of orfA and orfB. TM logo, . A strain was produced which lacks functional orfA, 〇rfB, and orfC' and contains a randomly binding Schizochytrium strain ATCC PTA-10212. This strain was identified by pLR95 containing the codon-optimized Schizochytrium strain ATCC PTA-10212 PFW (SEQ ID NO: i2〇) and the codon-optimized Schizochytrium strain ATCC PTA-10212 No.: 121) The pLR85 is transformed into a random combination with/to the household. The resulting recombinant strain lacks functional orfA, orfB, and orfc 153 201038734 as well as Schistosomiasis ATCC PTA-10212, PFX2' and PF/13 containing randomly bound Schizochytrium species. The cell lines were grown and analyzed for FAMEs as described in Example 7. The recombinant strain had a terpene content of 1.8% FAME, a DPAn-3 content of 1.8%, a DPAn-6 content of 2.3%, and a DHA content of 34.1%. Example 15 The orfA, orffl, and orfC genes from Schizochytrium sp. ATCC 20888 were colonized into a series of Duet vectors (Novagen). The Duet performance vector is a set of compatible plastids in which multiple target gene lines are selected and co-expressed by a T7-inducible promoter in E. coli. The Duet plastid pREZ91 contains the Schizochytrium strain ATCC 20888 orfA in pETDuet-1; the duet plastid pREZ96 contains the Schizochytrium sp. ATCC 20888 orffl in pCDFDuet-1; and the duet plastid pREZlOl contains the Schizochytrium sp. ATCC 20888 orfC is in pCOLADuet-1. The Duet plastids pREZ91, pREZ96, and pREZlOl, together with the desired helper gene Hetl plastid pJK737 (described in U.S. Patent No. 7,217,856, incorporated herein by reference in its entirety) Within the E. coli strain BLR (DE3), it contains an inducible T7 RNA polymerase gene. Once the cells were grown and IPTG was added, the DHA and DPAn-6 lines were produced according to the manufacturer's instructions (Novagen). Briefly, when the cells reached an optical density of about 0.5 at 600 nm, 1 mM IPTG was added for induction. The cells were grown in Luria broth for 3 hours at TC and harvested. These fatty acids were converted to methyl esters using standard techniques. The fatty acid morphology was determined using gas chromatography (GC_FID) with a flame ion detector. Fatty acid methyl ester 154 201038734 (FAME). Schizochytrium strain ATCC PTA-9695 corpse FW (SEQ ID NO: i) The gene line was selected into the expression vector pETDuet-1 to produce PREZ346. Duet plastid pREZ346 ( Containing Schizochytrium strains ATCC PTA-9695 E4/), pREZ96 (containing orffi), and pREZlOl (containing orfC) and pJK737 (containing Hetl) were transformed into E. coli strain 801 (0£3) The Schizophyllum sp. eight octopus (^? D-8-9695/>/^ gene line and the Schizochytrium strain ATCC 20888 orfB and orfC genes are co-expressed. Schizophyllum sp. ATCC PTA-9695, combined with fission The performance of the genus ATCC 20888 orffl and orfC supports the production of dha in E. coli under induction conditions. The transformed E. coli has a DHA content of 2.8% of FAME and a DPA n-6 content of 1.1%, DPA. The n-3 content was 0.6%, and the EPA content was 3.7%. Example 16 Codon-optimized rupture The strain ATCC PTA-10212 (sequence identification number: 120) was cloned into the expression vector pETDuet-Ι' to produce pLRIOO. Duet plastid pLRl00 (codon-preserving species ATCC containing codon optimization) PTA-10212 PE4/), PREZ96 (containing Schizochytrium strain ATCC 20888 orfB), and pREZlOl (containing Schizochytrium strain ATCC 20888 orfC) and pJK737 (containing Hetl) were transformed into E. coli strain BLR Within (DE3), see Example 15. Thraustochytrium sp. ATCC PTA-10212 Huoke 7 gene line and Schizochytrium strain ATcc 2〇888 orfB and orfC genes are co-expressed. Thraustochytrium strain ATCC PTA-10212 PiM/ 'combination with the performance of Schizochytrium strains ATCC 20888 orffl and 155 201038734 orfC supports the production of E. coli and EpA in E. coli under induction conditions. Example 17 Schizophrenia strain ATCC PTA-9695 (SEQ ID NO: 5) The gene line was selected into the expression vector pCOLADueH to produce pREZ326. Duet plastid pREZ326 (containing Schizochytrium strain ATCC PTA-9695 PFW), pREZ91 (containing Schizochytrium species ATCc) 20888 orfA) 'and pREZ96 (containing Schizochytrium strain ATCC 2〇888 〇rfB) and p JK737 (containing Hetl) was transformed into E. coli strain BLR (DE3). See Example 15. The Schizochytrium strain ATCC ΡΤΑ-9695, combined with the performance of the Schizochytrium strains ATCC 20888 orfA and orfB, supports the production of DHA in E. coli under induction conditions. The cell lines were grown and analyzed for FAMEs as described in Example 15. The transformed E. coli had a DHA content of 〇·3% FAME. Example 18 Codon-optimized Thraustochytrium sp. ATCC PTA-10212 (SEQ ID NO: 122) gene line was selected into the expression vector pCOLADuet-Ι to generate pREZ348°Duet plastid pREZ34 (with code Suboptimized Thraustochytrium strain ATCC PTA-10212 PE43), pREZ91 (containing Schizochyphora g ATCC 20888 orfA), and pREZ96 (containing Schizochytrium ATCC 20888 orfB) and PJK737 (containing Hetl) Together they were transformed into E. coli strain BLR (DE3). See Example 15. The Thraustochytrium strain ATCC PTA-l〇2l2, combined with the performance of Schizochytrium strains ATCC 20888 orfA and orfB, supports the production of DHA in the intestines under induction conditions 156 201038734. The cell lines were grown and analyzed for FAMEs as described in Example 15. The transformed E. coli had a DHA content of 29% FAME, and the DPAn-6 content was 0_4 〇/〇. Example 19 Schizochytrium strain ATCC PTA-9695 PE42 (SEQ ID NO: 3) The gene line was selected into the expression vector pCDFDuet-Ι, resulting in

pREZ330。Duet質體pREZ330 (含有裂殖壺菌種atcC PTA-9695 /7¾2)、pREZ326 (含有裂殖壺菌種 atcC PTA-9695户柯3)，以及pREZ91 (含有裂殖壺菌種ATCC 20888 orfA)係與pJK737 (含有Hetl) —起被轉形至大腸桿菌 函株BLR(DE3)内。參見實施例9。裂瘦壺菌種atcc PTA-9695尸和户，組合以裂殖壺菌種ATCc 2〇888 orfA的表現，支持誘導條件下之大腸桿菌内的1)11八的生 產。細胞係如實施例15中說明的方式生長且分析FAMEs。 該經轉形的大腸桿菌之DHA含量為0.8%的FAME，以及該 DPAn-6含量為0.2%。 實施例20 密碼子最佳化的破囊壺菌種ATCC PTA-10212 (序列辨識編號：121)基因係被選殖至該表現載體 pCDFDuet-Ι之内，產生pLR87<)Duet質體pLR87 (含有密碼 子最佳化的破囊壺菌種ATCC ΡΤΑ-10212 ΡΛ42)、pREZ348 (含有密碼子最佳化的破囊壺菌種ATCC pta_1〇212 户抑3) ’以及pREZ91 (含有裂殖壺菌種ATCC 20888 orfA)係 與pJK737 (含有Hetl)—起被轉形至大腸桿菌菌株BLr (DE3) 157 201038734 内。參見實施例15。密碼子最佳化的破囊壺菌種ATCC PTA-10212尸抑2和尸抑_3，組合以裂殖壺菌種ATCc 2〇888 orfA之表現，支㈣導條件下之切桿菌⑽DHA與低位 準之EPA的生產。細胞係如實施例15中說明的方式生長且 分析FAMEs。該經轉形的大腸桿菌2DHA含量為4 4%的 FAME ’ DPA n-6含# 為 1.1% ’以及EpA含量為〇 1%。 實施例21pREZ330. Duet plastid pREZ330 (containing Schizochytrium strain atcC PTA-9695 /73⁄42), pREZ326 (containing Schizochytrium strain atcC PTA-9695 keke 3), and pREZ91 (containing Schizochytrium strain ATCC 20888 orfA) It was transformed into E. coli BLR (DE3) together with pJK737 (containing Hetl). See Example 9. The scutellaria sinensis strain atcc PTA-9695 corpse and household, combined with the performance of the Schizochytrium strain ATCc 2〇888 orfA, supports the production of 1) 11 8 in E. coli under induction conditions. Cell lines were grown and analyzed for FAMEs as described in Example 15. The transformed E. coli had a DHA content of 0.8% FAME, and the DPAn-6 content was 0.2%. Example 20 Codon-optimized Thraustochytrium sp. ATCC PTA-10212 (SEQ ID NO: 121) gene line was selected into the expression vector pCDFDuet-Ι to generate pLR87<) Duet plastid pLR87 (containing Codon-optimized Thraustochytrium sp. ATCC ΡΤΑ-10212 ΡΛ42), pREZ348 (coccidial fungus strain ATCC pta_1〇212 containing 3) and pREZ91 (containing Schizochytrium species) ATCC 20888 orfA) was transformed into E. coli strain BLr (DE3) 157 201038734 with pJK737 (containing Hetl). See Example 15. Codon-optimized Thraustochytrium sp. ATCC PTA-10212 sin 2 and sin _3, combined with the performance of Schizochytrium strain ATCc 2〇888 orfA, cut (4) DHA and low position The production of EPA. The cell lines were grown and analyzed for FAMEs as described in Example 15. The transformed E. coli 2DHA content of 4% by weight of FAME ' DPA n-6 containing # was 1.1% ' and the EpA content was 〇 1%. Example 21

Duet質體pREZ346 (含有裂殖壺菌種八1(：€ PTA-9695 尸/^)、pREZ330 (含有裂殖壺菌種ATccptA-9695 尸E42)， 以及pREZ326 (含有裂殖壺菌種ATCC pTA_9695户尺係與 pJK737 (含有Hetl)—起被轉形至大腸桿菌菌株BLR (DE3) 内。參見實施例15。裂殖壺菌種ATCC pTA 9695户抑7、 PFJ2 ’以及尸E43的表現支持誘導條件下之大腸桿菌内的 DHA的生產。細胞係如實施例15中說明的方式生長且分析 FAMES。該經轉形的大腸桿菌之dha含量為〇 3。/〇的 FAME，以及EPA含量為0.3%。 實施例22Duet plastid pREZ346 (containing Schizochytrium strain VIII (: € PTA-9695 corpse / ^), pREZ330 (containing Schizochytrium strain ATccptA-9695 corpse E42), and pREZ326 (containing Schizochytrium strain ATCC pTA_9695 The pedigree was transformed into E. coli strain BLR (DE3) with pJK737 (containing Hetl). See Example 15. Schizochytrium sp. ATCC pTA 9695, PFJ2' and cadaveric E43 support expression induction Production of DHA in E. coli under conditions. The cell line was grown and analyzed for FAMES in the manner described in Example 15. The transduced E. coli had a dha content of 〇3./〇FAME, and an EPA content of 0.3. %. Example 22

Duet質體pLRIOO (含有密碼子最佳化的破囊壺菌種 ATCC ΡΤΑ-1〇212Ρ7^7)、ρυΐ87 (含有密碼子最佳化的破囊 壺菌種ATCC ΡΤΑ-10212 />柯2)，以及pREZ348 (含有密碼子 最佳化的破囊壺菌種ATCC ΡΤΑ-10212 /^3)係與pJK737 (含有Hetl)—起被轉形至大腸桿菌菌株BLR(DE3)内。參見 實施例15。密碼子最佳化的破囊壺菌種ATCC pta_1〇212 ’以及PE43之表現支持誘導條件下之大腸桿菌 158 201038734 内的DHA與ΕΡΑ的生產。 實施例23 ΟDuet plastid pLRIOO (codon-preserving strain ATCC ΡΤΑ-1〇212Ρ7^7 containing codon optimization), ρυΐ87 (coccidial bacterium containing ATCC ΡΤΑ-10212 /> Co2 ), and pREZ348 (coccidial fungus ATCC ΡΤΑ-10212 /^3 containing codon optimization) was transformed into E. coli strain BLR (DE3) with pJK737 (containing Hetl). See Example 15. The codon-optimized species of Thraustochytrium sp. ATCC pta_1〇212' and PE43 support the production of DHA and sputum in E. coli 158 201038734 under induction conditions. Example 23

Duet質體pREZ330 (含有裂殖壺菌種ATCC ΡΤΑ-9695 PFW)、pREZ91 (含有裂殖壺菌種ATCC 20888 orfA)，以及 pREZlOl (含有裂殖壺菌種ATCC 20888 orfC)係與pJK737 (含有Hetl)—起被轉形至大腸桿菌菌株BLR (DE3)内。參見 實施例15。裂瘦壺菌種ATCC ΡΤΑ-9695 /7^2，組合以裂殖 壺菌種ATCC 20888 orfA和orfC的表現，支持誘導條件下之 大腸桿囷内的DHA的生產。細胞係如實施例1 $中說明的方 式生長且分析FAMEs。該經轉形的大腸桿菌之DHA含量為 0.6%的FAME，以及DPAn-6含量為〇.3〇/0。 實施例24Duet plastid pREZ330 (containing Schizochytrium strain ATCC ΡΤΑ-9695 PFW), pREZ91 (containing Schizochytrium strain ATCC 20888 orfA), and pREZlOl (containing Schizochytrium strain ATCC 20888 orfC) and pJK737 (containing Hetl ) - was transformed into E. coli strain BLR (DE3). See Example 15. The A. serrata ATCC ΡΤΑ-9695 /7^2, combined with the performance of the Schizochytrium strains ATCC 20888 orfA and orfC, supports the production of DHA in the large intestines under induction conditions. Cell lines were grown and assayed for FAMEs as described in Example 1 $. The transformed E. coli had a DHA content of 0.6% FAME and a DPAn-6 content of 〇.3 〇/0. Example 24

Duet質體PLR87 (含有密碼子最佳化的破囊壺菌種 ATCC PTA-刪2觸2)、pREZ91 (含有裂殖壺菌種atcc 2_ 〇rfA)，以及pREZ101 (含有裂瘦壺菌種atcc 2〇888 orfC)係與P^37 (含有HetI) 一起被轉形至大腸桿菌菌株 BLR(DE3)内。參見實施例15。密碼子最佳化的破囊壺菌種 ATCC PTA-10212胸，組合以裂殖壺菌種ATCC 20888 orfA和orfC之表現，支持誘導條件下之大腸桿菌内的應 與低位準之EM的生產。細胞係如實施例i5中說_方式 生長且分析FAMES。該經轉形的大腸桿菌之腿含量為 1.7% 的 FAME，DPA n-6 含量為 0·9〇/〇， 0.1%。 以及EPA的含量為 實施例25 201038734Duet plastid PLR87 (coccidial bacterium containing ATCC PTA- deleted 2 touch 2), pREZ91 (containing Schizochytrium strain atcc 2_ 〇rfA), and pREZ101 (containing Schizochytrium species atcc) 2〇888 orfC) was transformed into E. coli strain BLR (DE3) together with P^37 (containing HetI). See Example 15. Codon-optimized Thraustochytrium sp. ATCC PTA-10212 thorax, combined with the performance of Schizochytrium strains ATCC 20888 orfA and orfC, supports the production of EM in low-level E. coli under induction conditions. The cell line was grown as described in Example i5 and analyzed for FAMES. The transformed E. coli had a leg content of 1.7% FAME and a DPA n-6 content of 0.19 〇/〇, 0.1%. And the content of EPA is Example 25 201038734

Duet質體pREZ346 (含有裂殖壺菌種atcc pta-9695 、pREZ330 (含有裂殖壺菌種ATCC PTA-9695尸柯乃， 以及pREZlOl (含有裂殖壺菌種ATCC 2〇888 orfC)係與 pJK737 (含有Hetl)—起被轉形至大腸桿菌菌株BLr (DE3) 内。參見實施例15。/>柯7與户抑2,組合以裂殖壺菌種ATCC 20888 orfC之表現，支持誘導條件下之大腸桿菌内的DHA 的生產。細胞係如實施例15中說明的方式生長且分析 FAMES。該經轉形的大腸桿菌之DHA含量為〇·3%的 FAME，DPAn-6含量為 0.1%，以及ερα含量為 〇.50/〇。 實施例26Duet plastid pREZ346 (containing Schizochytrium strain atcc pta-9695, pREZ330 (containing Schizochytrium strain ATCC PTA-9695 nectar, and pREZlOl (containing Schizochytrium ATCC 2〇888 orfC) and pJK737 (containing Hetl) - was transformed into E. coli strain BLr (DE3). See Example 15. /> Ke 7 and Hushen 2, combined with the performance of Schizochytrium strain ATCC 20888 orfC, support induction conditions Production of DHA in Escherichia coli. The cell line was grown and analyzed for FAMES in the manner described in Example 15. The transformed E. coli had a DHA content of 〇·3% FAME and a DPAn-6 content of 0.1%. And the ερα content is 〇.50/〇. Example 26

Duet質體pLRIOO (含有密碼子最佳化的破囊壺菌種 ATCC PTA-10212 PE4/)、pLR87 (含有密碼子最佳化的破囊 壺菌種ATCC PTA-10212 以及pREzi〇i (含有裂殖壺 菌種ATCC 20888 orfC)係與pJK737 (含有Hetl)—起被轉形 至大腸桿菌菌株BLR (DE3)内。參見實施例15。密碼子最佳 化的破囊壺菌種ATCC PTA-10212，組合以裂 殖壺菌種ATCC 20888 orfC之表現，支持誘導條件下之大腸 桿菌内的DHA與EPA的生產。 實施例27Duet plastid pLRIOO (codon-precipitating strain ATCC PTA-10212 PE4/) with codon optimization, pLR87 (coccidial bacterium containing ATCC PTA-10212 and pREzi〇i containing codon optimization) The bacterium strain ATCC 20888 orfC) was transformed into E. coli strain BLR (DE3) with pJK737 (containing Hetl). See Example 15. Codon-optimized Thraustochytrium ATCC PTA-10212 Combined with the performance of Schizochytrium strain ATCC 20888 orfC, it supports the production of DHA and EPA in E. coli under induction conditions.

Duet質體pREZ346 (含有裂殖壺菌種ATCC pta_9695 尸、pREZ% (含有裂殖壺菌種atCC 20888 orfB)，以及 pREZ326 (含有裂殖壺菌種ATCC PTA-9695 PE43)係與 pJK737 (含有Hetl)—起被轉形至大腸桿菌菌株BLR (DE3) 内。參見實施例15。裂殖壺菌種ATCC PTA-9695 與 160 201038734 PE43 ’組合以裂殖壺菌種ATCc 20888 orffi的表現，支持誘 導條件下之大腸桿菌内的DHA的生產。細胞係如實施例15 中說明的方式生長且分析FAMES。該經轉形的大腸桿菌之 DHA含量為0.1%的FAME，以及EPA含量為0.1%。 實施例28Duet plastid pREZ346 (containing Schizochytrium strain ATCC pta_9695 corpse, pREZ% (containing Schizochytrium strain atCC 20888 orfB), and pREZ326 (containing Schizochytrium strain ATCC PTA-9695 PE43) and pJK737 (containing Hetl ) - was transformed into E. coli strain BLR (DE3). See Example 15. Schizochytrium strains ATCC PTA-9695 and 160 201038734 PE43 'combined with the performance of Schizochytrium strain ATCc 20888 orffi, support induction Production of DHA in E. coli under conditions. The cell line was grown and analyzed for FAMES in the manner described in Example 15. The transformed E. coli had a DHA content of 0.1% FAME and an EPA content of 0.1%. Example 28

Duet質體pLRIOO (含有密碼子最佳化的破囊壺菌種 ATCC PTA-10212 />抑7)、pREZ96 (含有裂殖壺菌種ATCC 20888 orffi)，以及PREZ348 (含有密碼子最佳化的破囊壺菌 種ATCC PTA-10212 /^3)係與PJK737 (含有Hetl)-起被轉 形至大腸桿菌菌株BLR(DE3)内。參見實施例15。密碼子最 佳化的破囊壺菌種ATCC ΡΤΑ-1〇212 與PiMJ，組合以 裂殖壺菌種ATCC 20888 orfB之表現，支持誘導條件下之大 腸桿菌内的DHA與EPA的生產。 實施例29 裂殖壺菌種ATCC PTA-9695以及破囊壺菌種ATCC PTA-10212 中的Pfalp、Pfa2p，和Pfa3p PUFA合成酶活性係 分別地藉由標準的程序予以剔除。參見，例如，美國專利 案第7,217,856號，以其之整體併入本文中作為參考資料。 吉歐黴素™、潮黴素、殺稻瘟菌素(blasticidin)，或是 其他合適的抗性標誌係被插入至/基因（序列辨識編 號：1或序列辨識編號：68)的限制性位址之内，其被包含 於一質體内。插入該抗性標誌之後，該質體各別地被引入 至裂殖壺菌種ATCC PTA-9095或破囊壺菌種ATCC PTA-10212之内，其係藉由粒子轟擊法、電穿孔，或是其他 161 201038734 合適的轉形方法。發生同源性重組，產生突變體，其中天 然的基因係被抗吉歐黴素TM、潮黴素、殺稻瘟菌素， 或是其他合適的抗性標誌所取代或中斷的。轉形體係於含 有吉歐黴素™、潮黴素、殺稻瘟菌素，或是其他合適的篩 選劑，補充PUFAs的平盤上予以挑選。進一步檢查群落於 沒有PUFA補充的情況下生長的能力。基因體DNA係自對篩 選劑為抗性的以及無法於沒有PUFA補充的情況下生長之 群落單離。執行DNA的PCR和南方墨點分析以展現出Ρβί / 基因為删失或中斷的。 係藉由相似的程序予以剔除。發現需要PUFA補充 之生成的剔除突變體為缺少全長的户柯2。 係藉由相似的程序予以剔除。發現需要puFA補充 之生成的剔除突變體為缺少全長的。 本文中說明的各種各樣的態樣、具體例，以及選擇的 全體可以組合以任何及全部的變型。 此說月書中k及之全部的刊物、專利，以及專利申請 案係在相同程度上併人本文巾作為參考資料好似各個個 別的刊物、專利或專射請案被明確且個別地指示要被併 入本文中作為參考資料。 【圖式簡單說明】 弟1圖顯示出本發明沾則# ± — 个知月的裂殖亞菌種ATCC ΡΤΑ-9695 PUFA合成轉之基因架構； 第圖.‘、員tf出本發明的破囊壺菌種atcc ρτΑ ι〇2ΐ2 PUFA合成酶之基因架構； 162 201038734 第3圖顯示出本發明的裂殖壺菌種ATCC PTA-9695和 破囊壺菌種ATCC PTA-10212 PUFA合成酶以及來自裂殖壺 菌種ATCC 20888、破囊壺菌種ATCC 20892、金黄色破囊壺 菌，和SAM2179之合成酶之領域架構； 第4圖顯示出本發明裂殖壺菌種ATCC PTA-9695 Pfalp 胺基酸序列（序列辨識編號：2)和破囊壺菌種ATCC PTA-10212 Pfalp胺基酸序列（序列辨識編號：69)的排列， 其等具有該等之來自裂殖壺菌種ATCC 20888 (序列辨識編 號：54)和破囊壺菌種ATCC 20892 (序列辨識編號：56)的 OrfA序列以及來自金黃色破囊壺菌（序列辨識編號：55)的 OrfA序列； 第5圖顯示出排列本發明的裂殖壺菌種ATCC PTA-9695 Pfa2p胺基酸序列（序列辨識編號：4)和破囊壺菌 種ATCC PTA-10212 Pfa2p胺基酸序列（序列辨識編號：71) 的排列，其等具有該等之來自裂殖壺菌種ATCC 20888 (序 列辨識編號：57)和破囊壺菌種ATCC 20892 (序列辨識編 號：58)的OrfB序列以及來自金黄色破囊壺菌（序列辨識編 號：59)的OrfB序列； 第6圖顯示本發明的裂殖壺菌種ATCC PTA-9695 Pfa3p 胺基酸序列（序列辨識編號：6)以及破囊壺菌種ATCC PTA-10212 Pfa3p胺基酸序列（序列辨識編號：73)的排列， 其等具有來自裂殖壺菌種ATCC 20888 (序列辨識編號：61) 以及破囊壺菌種ATCC 20892 (序列辨識編號：60)的OrfC序 列； 163 201038734 第7圖顯示裂殖壺菌種ATCC PTA-9695 多核苷酸 序列（序列辨識編號：1); 第8圖顯示裂殖壺菌種ATCC PTA-9695 Pfalp胺基酸序 列（序列辨識編號：2); 第9圖顯示裂殖壺菌種ATCC ΡΤΑ-9695 多核苷酸 序列（序列辨識編號：3); 第1〇圖顯示裂殖壺菌種ATCC PTA-9695 Pfa2p胺基酸 序列(序列辨識編號：4); 第11圖顯示裂殖壺菌種ATCC PTA-9695 多核苷 酸序列（序列辨識編號：5); 第12圖顯示裂殖壺菌種ATCC PTA-9695 Pfa3p胺基酸 序列（序列辨識編號：6); 第13圖顯示破囊壺菌種ATCC PTA-10212 P/Mi多核苷 酸序列（序列辨識編號：68); 第14圖顯示破囊壺菌種ATCC PTA-10212 P抑/多核苷 酸序列（序列辨識編號：120)，其已經為了於裂殖壺菌内表 現而被密碼子最佳化； 第I5圖顯示破囊壺菌種ATCC PTA-10212 Pfalp胺基酸 序列(序列辨識編號：69); 第16圖顯示破囊壺菌種ATCC PTA-10212 7"柯2多核苷 酸序列（序列辨識編號：70); 第Π圖顯示破囊壺菌種ATCC PTA-10212 多核苷 酸序列（序列辨識編號：121)，其已經為了於裂殖壺菌内表 現而被密碼子最佳化； 164 201038734 第18圖顯示破囊壺菌種ATCC PTA-10212 Pfa2p胺基酸 序列（序列辨識編號：71); 第19圖顯示第13圖顯示破囊壺菌種ATCc PTA-10212 户E43多核苷酸序列（序列辨識編號：72); 第20圖顯示破囊壺菌種ATCc PTA_1〇2l2 多核苷 酸序列（序列辨識編號：122)，其已經為了於裂殖壺菌内表 現而被密碼子最佳化； 第21圖顯示破囊壺菌種ATCC PTA-10212 Pfa3p胺基酸 〇 序列(序列辨識編號：73); 第22圖顯示裂殖壺菌的密碼子使用表。 【主要元件符號說明】 - (無） 〇 165 201038734 <110〉 e id -—I i • s V s E e a s LD o k R r , f i R N / κ E ο E T s L ,H p K TCMR pill A R s z 表 列 序 <120> 多不飽和脂肪酸合成酶核酸分子及多肽，組成物，以及其製造方法與用途Duet plastid pLRIOO (codon-precursor strain ATCC PTA-10212 /> 7) with codon optimization, pREZ96 (containing Schizochytrium strain ATCC 20888 orffi), and PREZ348 (with codon optimization) The Thraustochytrium strain ATCC PTA-10212 /^3) was transformed into E. coli strain BLR (DE3) with PJK737 (containing Hetl). See Example 15. The codon-preserved strains of the genus Coccidiostats ATCC ΡΤΑ-1〇212 and PiMJ, combined with the performance of the Schizochytrium strain ATCC 20888 orfB, support the production of DHA and EPA in E. coli under induced conditions. Example 29 The Schiff's sp. ATCC PTA-9695 and the Pfalp, Pfa2p, and Pfa3p PUFA synthase activities in the Thraustochytrium ATCC PTA-10212 were individually removed by standard procedures. See, for example, U.S. Patent No. 7,217,856, the disclosure of which is incorporated herein by reference. GiomycinTM, hygromycin, blasticidin, or other suitable marker of resistance inserted into the restriction of the /gene (sequence ID: 1 or sequence ID: 68) Within the site, it is contained within a plastid. After insertion of the resistance marker, the plastid is separately introduced into the Schizochytrium sp. ATCC PTA-9095 or the Thraustochytrium sp. ATCC PTA-10212 by particle bombardment, electroporation, or Is the other 161 201038734 suitable transformation method. Homologous recombination occurs, producing mutants in which the natural gene line is replaced or interrupted by anti-GiomycinTM, hygromycin, blasticidin, or other suitable marker of resistance. The transformation system is selected on a flat plate containing chloramphenicolTM, hygromycin, blasticidin, or other suitable screening agent to supplement the PUFAs. The ability of the colonies to grow without PUFA supplementation was further examined. The genomic DNA is isolated from the colony that is resistant to the screening agent and cannot grow without PUFA supplementation. Perform DNA PCR and Southern blot analysis to show that the Ρβί / gene is censored or interrupted. It is eliminated by a similar procedure. The knockout mutants that were found to require PUFA supplementation were found to be lacking full length of the family. It is eliminated by a similar procedure. The knockout mutants that were found to require puFA supplementation were found to lack full length. The various aspects, specific examples, and alternatives described herein can be combined in any and all variations. All the publications, patents, and patent applications in the monthly book are in the same degree. The papers are used as reference materials. It is as if individual publications, patents, or special shots are clearly and individually indicated. It is incorporated herein by reference. [Simple description of the schema] Figure 1 shows the gene structure of the invention of the present invention: #±- 知 的 的 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 695 Cytosolic species atcc ρτΑ ι〇2ΐ2 gene architecture of PUFA synthase; 162 201038734 Figure 3 shows the Schizochytrium sp. ATCC PTA-9695 and the Thraustochytrium sp. ATCC PTA-10212 PUFA synthase and from Schistosomiasis ATCC 20888, Thraustochytrium ATCC 20892, Thraustochytrium aureum, and SAM2179 synthetase domain architecture; Figure 4 shows the Schizochytrium strain ATCC PTA-9695 Pfalp amine of the present invention Arrangement of the base acid sequence (SEQ ID NO: 2) and the Thraustochytrium strain ATCC PTA-10212 Pfalp amino acid sequence (SEQ ID NO: 69), which have the same from the Schizochytrium sp. ATCC 20888 ( Sequence identification number: 54) and OrfA sequence of Thraustochytrium sp. ATCC 20892 (SEQ ID NO: 56) and OrfA sequence from Thraustochytrium aureus (SEQ ID NO: 55); Figure 5 shows the alignment Invented Schizochytrium strain ATCC PTA-9695 Pfa2p amino acid sequence ( Sequence identification number: 4) and the arrangement of the Thraustochytrium sp. ATCC PTA-10212 Pfa2p amino acid sequence (SEQ ID NO: 71), which have the same from the Schizochytrium sp. ATCC 20888 (SEQ ID NO: 57) OrfB sequence of Thraustochytrium sp. ATCC 20892 (SEQ ID NO: 58) and OrfB sequence from Thraustochytrium aureus (SEQ ID NO: 59); Figure 6 shows Schizochytrium of the present invention ATCC PTA-9695 Pfa3p amino acid sequence (SEQ ID NO: 6) and the arrangement of the Thraustochytrium strain ATCC PTA-10212 Pfa3p amino acid sequence (SEQ ID NO: 73), which have from Schizochytrium ATCC 20888 (SEQ ID NO: 61) and OrfC sequence of Thraustochytrium sp. ATCC 20892 (SEQ ID NO: 60); 163 201038734 Figure 7 shows Schizochytrium sp. ATCC PTA-9695 polynucleotide sequence (sequence Identification number: 1); Figure 8 shows the Schizochytrium strain ATCC PTA-9695 Pfalp amino acid sequence (SEQ ID NO: 2); Figure 9 shows the Schizochytrium strain ATCC ΡΤΑ-9695 polynucleotide sequence ( Sequence identification number: 3); Figure 1 shows Schizochytrium ATCC PTA-9695 Pfa2p amino acid sequence (SEQ ID NO: 4); Figure 11 shows Schizochytrium sp. ATCC PTA-9695 polynucleotide sequence (SEQ ID NO: 5); Figure 12 shows Schizochytrium ATCC PTA-9695 Pfa3p amino acid sequence (SEQ ID NO: 6); Figure 13 shows the Thraustochytrium sp. ATCC PTA-10212 P/Mi polynucleotide sequence (SEQ ID NO: 68); Figure 14 shows Thraustochytrium sp. ATCC PTA-10212 P/polynucleotide sequence (SEQ ID NO: 120), which has been codon optimized for expression in Schizochytrium; Figure I5 shows Thraustochytrium ATCC PTA-10212 Pfalp amino acid sequence (SEQ ID NO: 69); Figure 16 shows the Thraustochytrium strain ATCC PTA-10212 7 " Ke 2 polynucleotide sequence (SEQ ID NO: 70); The Thraustochytrium sp. ATCC PTA-10212 polynucleotide sequence (SEQ ID NO: 121), which has been codon-optimized for expression in Schizochytrium; 164 201038734 Figure 18 shows Thraustochytrium ATCC PTA-10212 Pfa2p amino acid sequence (SEQ ID NO: 71); Figure 19 shows Figure 13 shows Thraustochytrium strain ATCc PTA-10212 Family E43 polynucleotide sequence (SEQ ID NO: 72); Figure 20 shows the Thraustochytrium strain ATCc PTA_1〇2l2 polynucleotide sequence (SEQ ID NO: 122), which has Codon optimized for expression in Schizochytrium; Figure 21 shows the sequence of Thraustochytrium ATCC PTA-10212 Pfa3p amino acid hydrazide (SEQ ID NO: 73); Figure 22 shows Schizoter The codon usage table for the bacteria. [Description of main component symbols] - (none) 〇165 201038734 <110> e id -—I i • s V s E eas LD ok R r , fi RN / κ E ο ET s L , H p K TCMR pill AR Sz table order <120> polyunsaturated fatty acid synthase nucleic acid molecule and polypeptide, composition, and preparation method and use thereof

<130> 2715.025TW02/JUK/SAS <140> 待讓渡 <141> 2010-03-19 <150> 61/296,460 <151> 2010-01-19 <150> 61/161,742 <151» 2009-03-19 <160> <170> <210> <211> <212> <213> 〇 127<130> 2715.025TW02/JUK/SAS <140> To be transferred<141> 2010-03-19 <150> 61/296,460 <151> 2010-01-19 <150> 61/161,742 <;151» 2009-03-19 <160><170><210><211><212><213> 〇127

Patentln 版本 3_ 7824Patentln version 3_ 7824

DNA 裂殖壺菌物種 〇 <400> 1 atggatactc gcatcgcgat cgcgagagct gggaggcgat cgcgtggacg tgacggccta aagcgcggcg ggttcatccc ttccagatgg aggactcgga ctgacggacg ccaacatccc ggcatcggcg gcggccagaa gtggacaagg tgctgcgcaa aagtacaagg cgagtttccc gtcacggcgg ggcgctgctg gcggcctgcg cgtcgtcgct gactgcgatg cgatgatcgc gccttctcca agacgcccgt accaaaggca tgctcatcgg gccgtgcgcg acggcgacac ggcaaggcgg cgggcatcta gcctacgccc gcgccaatgt ggtacgccgg tgggcgacaa ttttctgcca acggtggcgg tcgcagatcg ggcacctcaa gcgctcaagc acaagacgct gacgggaccc cgatccagca cgtggggatg ccgcgatggg ctacaacccg ggagtacgac cgccaaccag ggcgttctcg ggcgagccac gatgggcctg cgagtggcgc caataccttc gatcgcggtc gggtgccacc gttttccacg cgagggctcg cgtgcacgcc cacgccgaca cgacccggcc gatcgagctg cgcggaggaa ggcggtggcc gccgcagacg gtcgccgctg tcggcgatcc ctggattgcc gagaagacga ttcgacgcgc acgatctcgc agcggtaaga gagttctact ccggaggaag ctcgactctt aacatggagg aaagtggcga tgcacggaca gacccgagcg gcgatgctcg gtcatcaagg atctcgggcc actgtgacgc acggcgctga gcagagcagg gggctggccg atcaacgtcg tacgtcaaca tgccgagcgg tgagcgatct ccaaggacaa gtgagttcgg tgctcaaggt agaacatcgg cgcggctcaa acgtggcggc tccccgggtt gcatgaactg tcgaggagct actcgatcgg tcaaggcgta tgctgaagcg ggtgcgcgtc aggaggaggc tggtggaggg gcaacctctt tggcggtggg ggctggtcaa acaagccgcc cgatgaaccg ggagaacgtg gccggcggac gatctactgc gctcaacatg gaaggaggcg ctgcgtgctg ctacgtggtc ggcggtggac cctgggcaac cgtcgtggac gctctacggc gatgtacatg cgacgccgcc ctacgcggac ctcgagcgac cctgcgccgc ccacggcacg ctccaaggcg cagcatcaag ggtggtgctg gtcgctggtg cccctggttc 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1 201038734 acgcccgtag gggtgccgcg ccgcgccggc gtgtcgtcgt ttgggtttgg cggtgccaac 1380 taccacgccg tgctggagga gtttgagccc gagcacgaga gcgcgtaccg gtacaacaac 1440 ctgccgcagg tggcgctgct gcacgcgggg gacgtcgcga ccttggcggc gacggttcgc 1500 gccaagctgg cgctggccac cgccgagcag gaagaggcgc gtgtggtgaa gaacgcggac 1560 tacatcgcgt accaccggtt cctggacgag tgcaagttgc gcggcgctgt gccgcaggcg 1620 cacgcgcggg tgggactgct cgtacgggac ctgagctcgc tcatcgccgt gctcgaggcc 1680 gctgccgcca agctcgcggg cgaagagagc gcgacggagt ggacggtcag cgttgctacg 17 40 ggcgaggcgg ccttccgcgt gcgcggtgtg gctacggagg ccaacgtggc ggcgctgttc 1800 tcgggccagg gcgcgcagta cacgcacatg ttcagcgacg tggcgatgaa ctggcccccg 1860 ttccgcgaga gcgtcgccgc catggaccgc gcccagcgcg agcgcttcgg gcggcctgcc 1920 aagcgcgtga gcagcgtgct gtacccgcgc aagccgtacg gcgacgaacc gcggcaggac 1980 cacaaggaga tctcgcaaac gcgctactcg cagcccgcaa cgctcgcgtg ctcggtcggc 2040 gcctttgaca tcttcaaagc ggcgggactg gcgccgagct ttgcggcggg ccactcgctg 2100 ggcgagtttg cggcgctcta cgcggccggg tcgctcgatc gcgacgccgt cttcgacctg 2160 gtctgcgcgc gcgccaaggc catgagcgac ttcacggccc aggccagcag cagcggtggc 2220 gccatggcgg ccgtgattgg cgccaaggcg gaccagctct cgctgggtgg cgcgcccgac 2280 gtgtggctcg ccaacagcaa ctcgccctcg cagaccgtga tcacgggaac cgccgaagca 2340 gtggctgcgg cctctgacaa gttgcgctgc agcggcaact tccgcgtcgt gcctctggcc 2400 tgcgaggcgg ccttccactc gccgcacatg cgcggcgcgg agcagacgtt tgcgtcggcg 2460 ctcgcgcagg cgcccgtgtc ggcaccggcg gctgctcggt tctactctaa cgtgacgggg 2520 ggcgccgcgg taacctcgcc cgcggacgtc aaaacgaacc tgggcaagca catgacgagc 2580 cctgtgcagt tcgtgcagca ggtgcgagcc atgcacgcgg cgggcgcgcg tgtgtttgtg 2640 gagtttgggc ccaagcaggt cctgtcgcgc ctcgtcaagg agacccttgg cgaggccggc 2700 gacgtggtca cggtcgccgt caacccagac tcggccaagg acagcgacac gcagctgcgc 2760 caggcggcgc tcacgttggc ggtcgccggc gtgccgctca aggactttga ccgctggcag 2820 ctgccggatg ccacgcgcct cgagcctgtc aagaagaaga agaccacgtt gcggctctcg 2880 gcagccacct acgtctccgc caagacgttg cgccagcgcg aggccgtgct caacgacggc 2940 tacactgtca gtggtgccac ggcggtagtc aaggaagtgg acacggccaa cgaggagcgt 3000 ctcgtccgcc aagcccagga tctccagcgc cagctcgcgg aggcctcgac ggcagcccag 3060 gcggcgcagt ccaaggtcgc ggagctcgag cgcacgatcc aggacttgga gcgcaaggtg 3120 cagcagcagc agcaagagaa gggtgagaac tcagacagca acgctgccgc cgaagtgctg 3180 cggcgccaca aggagctgct ccagcgcatg ctgcaggact gtgacgagca ggcagtgccc 3240 gtagccacgg tggttccgac acctacgtcc tccccgacgc ctacatcctc acccgtatcc 3300 ggcaacagca agagcactcg tggcagtgct gatctgcaag cgctgctggc caaggcggag 3360 actgtggtga tggctgtgct ggctgccaag actggctacg aggccgacat ggttgaggcg 3420 gacatggacc tggaggccga gctcggcatc gactcgatca agcgcgtgga gatcctttcc 3480 gaggtgcagg gccagctggg cgtcgaggcc aaggacgtgg atgcgctgag ccgcacgcgc 3540DNA Schizochytrium species square < 400 > 1 atggatactc gcatcgcgat cgcgagagct gggaggcgat cgcgtggacg tgacggccta aagcgcggcg ggttcatccc ttccagatgg aggactcgga ctgacggacg ccaacatccc ggcatcggcg gcggccagaa gtggacaagg tgctgcgcaa aagtacaagg cgagtttccc gtcacggcgg ggcgctgctg gcggcctgcg cgtcgtcgct gactgcgatg cgatgatcgc gccttctcca agacgcccgt accaaaggca tgctcatcgg gccgtgcgcg acggcgacac ggcaaggcgg cgggcatcta gcctacgccc gcgccaatgt ggtacgccgg tgggcgacaa ttttctgcca acggtggcgg tcgcagatcg ggcacctcaa gcgctcaagc acaagacgct gacgggaccc cgatccagca cgtggggatg ccgcgatggg ctacaacccg ggagtacgac cgccaaccag ggcgttctcg ggcgagccac gatgggcctg cgagtggcgc caataccttc gatcgcggtc gggtgccacc gttttccacg cgagggctcg cgtgcacgcc cacgccgaca cgacccggcc gatcgagctg cgcggaggaa ggcggtggcc gccgcagacg gtcgccgctg tcggcgatcc ctggattgcc gagaagacga ttcgacgcgc acgatctcgc agcggtaaga gagttctact ccggaggaag ctcgactctt aacatggagg aaagtggcga tgcacggaca gacccgagcg gcgatgctcg gtcatcaagg atctcgggcc actgtgacgc acggcgctga gcagagcagg gggctggccg atcaacgtcg ta cgtcaaca tgccgagcgg tgagcgatct ccaaggacaa gtgagttcgg tgctcaaggt agaacatcgg cgcggctcaa acgtggcggc tccccgggtt gcatgaactg tcgaggagct actcgatcgg tcaaggcgta tgctgaagcg ggtgcgcgtc aggaggaggc tggtggaggg gcaacctctt tggcggtggg ggctggtcaa acaagccgcc cgatgaaccg ggagaacgtg gccggcggac gatctactgc gctcaacatg gaaggaggcg ctgcgtgctg ctacgtggtc ggcggtggac cctgggcaac cgtcgtggac gctctacggc gatgtacatg cgacgccgcc ctacgcggac ctcgagcgac cctgcgccgc ccacggcacg ctccaaggcg cagcatcaag ggtggtgctg gtcgctggtg cccctggttc 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1 201038734 acgcccgtag gggtgccgcg ccgcgccggc gtgtcgtcgt ttgggtttgg cggtgccaac 1380 taccacgccg tgctggagga gtttgagccc gagcacgaga gcgcgtaccg gtacaacaac 1440 ctgccgcagg tggcgctgct gcacgcgggg gacgtcgcga ccttggcggc gacggttcgc 1500 gccaagctgg cgctggccac cgccgagcag gaagaggcgc gtgtggtgaa gaacgcggac 1560 tacatcgcgt accaccggtt cctggacgag Tgcaagttgc gcggcgctgt gccgcaggcg 1620 cacgcgcggg tgggactgct cgtacgggac ctgagctcg c tcatcgccgt gctcgaggcc 1680 gctgccgcca agctcgcggg cgaagagagc gcgacggagt ggacggtcag cgttgctacg 17 40 ggcgaggcgg ccttccgcgt gcgcggtgtg gctacggagg ccaacgtggc ggcgctgttc 1800 tcgggccagg gcgcgcagta cacgcacatg ttcagcgacg tggcgatgaa ctggcccccg 1860 ttccgcgaga gcgtcgccgc catggaccgc gcccagcgcg agcgcttcgg gcggcctgcc 1920 aagcgcgtga gcagcgtgct gtacccgcgc aagccgtacg gcgacgaacc gcggcaggac 1980 cacaaggaga tctcgcaaac gcgctactcg cagcccgcaa cgctcgcgtg ctcggtcggc 2040 gcctttgaca tcttcaaagc ggcgggactg gcgccgagct ttgcggcggg ccactcgctg 2100 ggcgagtttg cggcgctcta cgcggccggg tcgctcgatc gcgacgccgt cttcgacctg 2160 gtctgcgcgc gcgccaaggc catgagcgac ttcacggccc aggccagcag cagcggtggc 2220 gccatggcgg ccgtgattgg cgccaaggcg gaccagctct cgctgggtgg cgcgcccgac 2280 gtgtggctcg ccaacagcaa ctcgccctcg cagaccgtga tcacgggaac cgccgaagca 2340 gtggctgcgg cctctgacaa gttgcgctgc agcggcaact tccgcgtcgt gcctctggcc 2400 tgcgaggcgg ccttccactc gccgcacatg cgcggcgcgg agcagacgtt tgcgtcggcg 2460 ctcgcgcagg cgcccgtgtc ggcaccggcg gctgctcggt tctactctaa cgtgacgggg 2520 ggcgccgcgg taacctcgcc cgcggacgtc aaaacgaacc tgggcaagca catgacgagc 2580 cctgtgcagt tcgtgcagca ggtgcgagcc atgcacgcgg cgggcgcgcg tgtgtttgtg 2640 gagtttgggc ccaagcaggt cctgtcgcgc ctcgtcaagg agacccttgg cgaggccggc 2700 gacgtggtca cggtcgccgt caacccagac tcggccaagg acagcgacac gcagctgcgc 2760 caggcggcgc tcacgttggc ggtcgccggc gtgccgctca aggactttga ccgctggcag 2820 ctgccggatg ccacgcgcct cgagcctgtc aagaagaaga agaccacgtt gcggctctcg 2880 gcagccacct acgtctccgc caagacgttg cgccagcgcg aggccgtgct caacgacggc 2940 tacactgtca gtggtgccac ggcggtagtc aaggaagtgg acacggccaa cgaggagcgt 3000 ctcgtccgcc aagcccagga tctccagcgc cagctcgcgg aggcctcgac ggcagcccag 3060 gcggcgcagt ccaaggtcgc ggagctcgag cgcacgatcc aggacttgga gcgcaaggtg 3120 cagcagcagc agcaagagaa gggtgagaac tcagacagca acgctgccgc cgaagtgctg 3180 cggcgccaca aggagctgct ccagcgcatg ctgcaggact gtgacgagca ggcagtgccc 3240 gtagccacgg tggttccgac acctacgtcc tccccgacgc ctacatcctc acccgtatcc 3300 ggcaacagca agagcactcg tgg Cagtgct gatctgcaag cgctgctggc caaggcggag 3360 actgtggtga tggctgtgct ggctgccaag actggctacg aggccgacat ggttgaggcg 3420 gacatggacc tggaggccga gctcggcatc gactcgatca agcgcgtgga gatcctttcc 3480 gaggtgcagg gccagctggg cgtcgaggcc aaggacgtgg atgcgctgag ccgcacgcgc 3540

2 2010387342 201038734

acggtcggtg gctcctgcgg gcgccttctg ctggcggcca gagctcggca ggcgtcgagg gatgccatga ccttcggcgc gatctgcaag actggctacg gactcgatca aaggacgtgg gcggaaatcg gctgcttctg ctgctggcca gccgacatgg cgcgtggaga gcgctgagcc gctgcctctg gctccgactc gcggagactg gaggcggaca ctctcggagg acgcgcacgg ggtagtgctc gcgactgcgc gctgtgctgg gaggccgagc cagctgggcg gtggtggacg ccttcggcgc acctttcccg gctcgccctg ctcggcgatc accacgcaca acgtccgtcg gacgtgcaag tcagaacaga aggttgtgga ttccttcggc ctgatctgca agactggcta tcgactcgat ccaaggacgt aggcggaaat ccgctgcttc cgctgctgtc aggccgacat agcgcgtgga atgcgctgag tggctgcctc cagctccgac aggcggagac tcgaggcgga tcctttccga gcacgcgcac ctggtagtgc ccgctgcttc tggtgatggc tggacctgga tgcagggcca tcggtgaggt ctgctgctgc cttctgctga cggccaagac tcggcatcga tcgaggccaa ccatgaaggc ccgcgcttct tgataacgac tagtcgtggt gtgcggtgct agttggtgac aggcgcagtt cgcagctcgg tcgagggcgg cgccatgaag gcccgctgct agcgctgctg cgaggccgac caagcgcgtg ggatgcgctg cgtggctgcc tgcagctccg caaggcggag ggtcgaggcg gatcctctcg ccgcacgcgc tggtggtagt tcccgcggct tgtggtgatg catggacctg ggtgcagggc ggtcggtgag tcctgctcct gactgcgcct tgtgctggcg ggccgagctc gctgggcgtc tgtggatgcc tgttccttcg tctgcaagcg tggctacgag ctcgatcaag ggacgtagat ggagatcgtg tccaacgctg cctgccgctt ggatgatgga gctgcaggtg cgtagcagac cggcaaggtg ctgggcgctg tcgcaccttt gcggagatcg tctgcagctc tccaaggcgg atggtcgagg gagatcctct agccgcacgc tctgctggta actcccgctg acggtggtga gacatggacc gaggtgcagg acggtcggtg gctcctgctc gcgacagcgc gctgtgctgg gaggccgagc cagctgggcg gttgtggatg gctgttcctt tctgctgatc gccaagactg ggcatcgact gaggccaagg atgaaggcgg gcgcccgctg ctgctgtcca gccgacatgg cgcgtggaga gcgctgagcc gctgcctctg tttggttccg cctgcagagc tctgcactca cagtcttcct cgctctgaag ggtggctttg ctcgcggcca ttcgtggccg tggctgcctc cgactcccgc agactgtggt cggacatgga cggaggtgca gcacggtcgg gtgctcctgc cttcgactgc tggctgtgct tggaggccga gccagctggg aggttgtgga ctgcggttcc cttctgctga cggccaagac tcggcatcga tcgaggccaa ccatgaaggc cggcgcccgc tgcaagcgct gctacgaggc cgatcaagcg acgtggatgc aaatcgtggc cttctgcagc aggcggagac tcgaggcgga tcctttccga gcacgcgcac gtggtagtgc agtgcgagga ttgtgctggc cctcgtcgct ctgcctgctc cggcgctaca tgttccagtt agcacctcaa tcgcgcggct tggtggtagt tgcttcgact gatggctgtg cctggaggcc gggccagctg tgaggttgtg tcctgctgtt gccttctgct ggcggccaag gctcggcatc cgtcgaggcc tgccatgaag ttcggcgccc tctgcaagcg tggctacgag ctcgatcaag ggacgtagat ggagatcgtg tgcttctgca gctgtccaag cgacatggtc cgtggagatc gctgagccgc tgcctctggt tccgactcct tgtggtgatg catggacctg ggtgcagggc ggtcggtgaa tcctgctgct cctgtctctg cgagggcggc ggtgtcctcg gccgcgctcg ggcggcgctc cggcgacgac aacttcgctg cgacggccag 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 3 201038734 ctggggctct ccggcaagtc gacgaccgct accgttgatc tctcccgcgc gcagcagggc 5880 agcgtgttcg gcctgtgcaa gacactcgac ctggagtggc ccgctgtctt ctgccgcgga 5940 atcgacctgg ccgccgacct cgacgccgca caggccgcgc ggtgcctgct gggcgagctg 6000 tcagaccccg acgtggccgt gcgcgagtct ggttactccg cctcgggcca gcgctgcacg 6060 acaactacga agtcgctgac tacgggcaag ccgcaccagc cgatctcctc gtcggacctc 6120 tttctggtgt cgggcggcgc gcgcggcatc accccgctgt gcgtgcgcga gctggcgcag 6180 cgcgtgggcg gcggcacgta cgtgctcatc ggccgctcgg agctgcccac gacggagcct 6240 gcctgggcgg tcggcgtgga gtctggcaag ccgctggaga aggccgcgct ggcgttcctg 6300 aaggcggagt ttgcagcggg ccgcggggcc aagccgacgc cgatgctgca caagaagctc 6360 gtgggcgccg tggtcggagc gcgcgaggtg cgagcctcgc tcgccgagat cactgcacag 6420 ggcgccacgg ctgtgtacga gtcgtgcgac gtgagctctg ccgccaaggt gcgtgagatg 6480 gtagagcgcg tgcagcagca gggcgggcgg cgcgtgtcgg gcgtgttcca cgcgtcgggc 6540 gtgctgcgcg acaagctcgt ggagaacaag tcgctggcgg acttcagcgc cgtgtacgac 6600 accaaggtgg gcggcctcat caacctgctg gcctgcgtgg acctggcgca gctgcgtcac 6660 ctcgtgctct tcagctcgct cgcgggcttc cacggcaacg tcgggcagtc ggactacgca 6720 atggccaacg aggcgctcaa caagctggcg gcgcacctgt cggcggtgca cccgcagctg 6780 tgcgcgcgct cgatctgctt cggaccgtgg gacggcggca tggtgacccc cgcgctcaag 6840 gccaacttca tccgcatggg catccagatc atcccgcgcc aaggcggcgc gcagaccgtc 6900 gccaacatgc tcgtcagtag ctcccccggt cagctgctcg tgggcaactg gggcgtgcca 6960 cccgtcgtgc cgagtgccac cgagcacacc gtgctgcaga cgctccgcca gagcgacaac 7020 cccttcctcg actcgcacgt gatccagggc cgccgcgtgc tgcccatgac cctggccgtg 7080 ggctacatgg cgcaccaggc gcagagcatc tacgcgggcc accagctgtg ggccgtcgag 7140 gacgcccagc tcttcaaggg catcgccatc gacaatggcg ccgacgtgcc cgtgcgcgtg 7200 gagctgtcgc gccgcaagga ggagcaggag gacgccggca aggtcaaggt caaggtgcag 7260 gtgctgctca aatcgcaggt caacggcaag tcggtgcccg cgtacaaggc gaccgtcgtg 7320 ctgtcccctg cgccgcgccc cagcgtcatc acgcgtgact tcgacctcac cccggacccg 7380 gcctgcacgg agcacgacct ctacgacggc aagacgctct tccacggcaa ggccttccag 7440 ggcatcgagc aggtgctctc ggcgacgccc aagcagctca ccgccaagtg ccgcaatttg 7500 cccctcacgc ccgagcagcg cggccagttc gtcgttaacc tcagccagca ggacccgttc 7560 caggcggaca ttgcgttcca ggcgatgctc gtctgggcgc gcatgctgcg ccaatcggcg 7620 gccctgccca acaactgcga gcgcttcgac ttttacaagc cgatggcccc gggcgccacc 7680 tactacacgt cggtcaagct ggcctcggcc tcacccttgg tggactctgt gtgcaagtgc 77 40 accgtggcga tgcacgatga gcaaggtgag gtgtactttt ctgctcgtgc cagcgtcgtc 7800 ctcaacaaga ccctcacgta ctaa 7824acggtcggtg gctcctgcgg gcgccttctg ctggcggcca gagctcggca ggcgtcgagg gatgccatga ccttcggcgc gatctgcaag actggctacg gactcgatca aaggacgtgg gcggaaatcg gctgcttctg ctgctggcca gccgacatgg cgcgtggaga gcgctgagcc gctgcctctg gctccgactc gcggagactg gaggcggaca ctctcggagg acgcgcacgg ggtagtgctc gcgactgcgc gctgtgctgg gaggccgagc cagctgggcg gtggtggacg ccttcggcgc acctttcccg gctcgccctg ctcggcgatc accacgcaca acgtccgtcg gacgtgcaag tcagaacaga aggttgtgga ttccttcggc ctgatctgca agactggcta tcgactcgat ccaaggacgt aggcggaaat ccgctgcttc cgctgctgtc aggccgacat agcgcgtgga atgcgctgag tggctgcctc cagctccgac aggcggagac tcgaggcgga tcctttccga gcacgcgcac ctggtagtgc ccgctgcttc tggtgatggc tggacctgga tgcagggcca tcggtgaggt ctgctgctgc cttctgctga cggccaagac tcggcatcga tcgaggccaa ccatgaaggc ccgcgcttct tgataacgac tagtcgtggt gtgcggtgct agttggtgac aggcgcagtt cgcagctcgg tcgagggcgg cgccatgaag gcccgctgct agcgctgctg cgaggccgac caagcgcgtg ggatgcgctg cgtggctgcc tgcagctccg caaggcggag ggtcgaggcg gatcctctcg ccgcacgcgc tggtggtagt tcccgcggct tgtggtgatg catggacctg ggtgcagggc ggtcggtgag tcctgctcct gactgcgcct tgtgctggcg ggccgagctc gctgggcgtc tgtggatgcc tgttccttcg tctgcaagcg tggctacgag ctcgatcaag ggacgtagat ggagatcgtg tccaacgctg cctgccgctt ggatgatgga gctgcaggtg cgtagcagac cggcaaggtg ctgggcgctg tcgcaccttt gcggagatcg tctgcagctc tccaaggcgg atggtcgagg gagatcctct agccgcacgc tctgctggta actcccgctg acggtggtga gacatggacc gaggtgcagg acggtcggtg gctcctgctc gcgacagcgc gctgtgctgg gaggccgagc cagctgggcg gttgtggatg gctgttcctt tctgctgatc gccaagactg ggcatcgact gaggccaagg atgaaggcgg gcgcccgctg ctgctgtcca gccgacatgg cgcgtggaga gcgctgagcc gctgcctctg tttggttccg cctgcagagc tctgcactca cagtcttcct cgctctgaag ggtggctttg ctcgcggcca ttcgtggccg tggctgcctc cgactcccgc agactgtggt cggacatgga cggaggtgca gcacggtcgg gtgctcctgc cttcgactgc tggctgtgct tggaggccga gccagctggg aggttgtgga ctgcggttcc cttctgctga cggccaagac tcggcatcga tcgaggccaa ccatgaaggc cggcgcccgc tgcaagcgct gctacgaggc cgatcaagcg acgtggatgc aaatcgtggc cttctgcagc aggcggagac tcgaggcgga tcctttccga gcacgcgcac gtggtagtg c agtgcgagga ttgtgctggc cctcgtcgct ctgcctgctc cggcgctaca tgttccagtt agcacctcaa tcgcgcggct tggtggtagt tgcttcgact gatggctgtg cctggaggcc gggccagctg tgaggttgtg tcctgctgtt gccttctgct ggcggccaag gctcggcatc cgtcgaggcc tgccatgaag ttcggcgccc tctgcaagcg tggctacgag ctcgatcaag ggacgtagat ggagatcgtg tgcttctgca gctgtccaag cgacatggtc cgtggagatc gctgagccgc tgcctctggt tccgactcct tgtggtgatg catggacctg ggtgcagggc ggtcggtgaa tcctgctgct cctgtctctg cgagggcggc ggtgtcctcg gccgcgctcg ggcggcgctc cggcgacgac aacttcgctg cgacggccag 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 3 201038734 ctggggctct ccggcaagtc gacgaccgct accgttgatc tctcccgcgc gcagcagggc 5880 agcgtgttcg gcctgtgcaa gacactcgac ctggagtggc ccgctgtctt ctgccgcgga 5940 atcgacctgg ccgccgacct cgacgccgca caggccgcgc ggtgcctgct gggcgagctg 6000 tcagaccccg acgtggccgt gcgcgagtct ggttactccg cctcgggcca gcgctgcacg 6060 ac aactacga agtcgctgac tacgggcaag ccgcaccagc cgatctcctc gtcggacctc 6120 tttctggtgt cgggcggcgc gcgcggcatc accccgctgt gcgtgcgcga gctggcgcag 6180 cgcgtgggcg gcggcacgta cgtgctcatc ggccgctcgg agctgcccac gacggagcct 6240 gcctgggcgg tcggcgtgga gtctggcaag ccgctggaga aggccgcgct ggcgttcctg 6300 aaggcggagt ttgcagcggg ccgcggggcc aagccgacgc cgatgctgca caagaagctc 6360 gtgggcgccg tggtcggagc gcgcgaggtg cgagcctcgc tcgccgagat cactgcacag 6420 ggcgccacgg ctgtgtacga gtcgtgcgac gtgagctctg ccgccaaggt gcgtgagatg 6480 gtagagcgcg tgcagcagca gggcgggcgg cgcgtgtcgg gcgtgttcca cgcgtcgggc 6540 gtgctgcgcg acaagctcgt ggagaacaag tcgctggcgg acttcagcgc cgtgtacgac 6600 accaaggtgg gcggcctcat caacctgctg gcctgcgtgg acctggcgca gctgcgtcac 6660 ctcgtgctct tcagctcgct cgcgggcttc cacggcaacg tcgggcagtc ggactacgca 6720 atggccaacg aggcgctcaa caagctggcg gcgcacctgt cggcggtgca cccgcagctg 6780 tgcgcgcgct cgatctgctt cggaccgtgg gacggcggca tggtgacccc cgcgctcaag 6840 gccaacttca tccgcatggg catccagatc atcccgcgcc aaggcggcgc gcagaccgtc 6900 gccaacatgc tcgtcagtag ctcccccggt cagctgctcg tgggcaactg gggcgtgcca 6960 cccgtcgtgc cgagtgccac cgagcacacc gtgctgcaga cgctccgcca gagcgacaac 7020 cccttcctcg actcgcacgt gatccagggc cgccgcgtgc tgcccatgac cctggccgtg 7080 ggctacatgg cgcaccaggc gcagagcatc tacgcgggcc accagctgtg ggccgtcgag 7140 gacgcccagc tcttcaaggg catcgccatc gacaatggcg ccgacgtgcc cgtgcgcgtg 7200 gagctgtcgc gccgcaagga ggagcaggag gacgccggca aggtcaaggt caaggtgcag 7260 gtgctgctca aatcgcaggt caacggcaag tcggtgcccg cgtacaaggc gaccgtcgtg 7320 ctgtcccctg cgccgcgccc cagcgtcatc acgcgtgact tcgacctcac cccggacccg 7380 gcctgcacgg agcacgacct ctacgacggc aagacgctct tccacggcaa ggccttccag 7440 ggcatcgagc aggtgctctc ggcgacgccc aagcagctca ccgccaagtg ccgcaatttg 7500 cccctcacgc ccgagcagcg cggccagttc gtcgttaacc tcagccagca ggacccgttc 7560 caggcggaca ttgcgttcca ggcgatgctc gtctgggcgc gcatgctgcg ccaatcggcg 7620 gccctgccca acaactgcga gcgcttcgac ttttacaagc cgatggcccc gggcgccacc 7680 tactacacgt cggtcaagct ggcctcggcc tcacccttgg tggactctgt gtg Caagtgc 77 40 accgtggcga tgcacgatga gcaaggtgag gtgtactttt ctgctcgtgc cagcgtcgtc 7800 ctcaacaaga ccctcacgta ctaa 7824

<210> 2 <211> 2607 <212> PRT <213> 裂殖壺菌物種 4 201038734 <400> 2<210> 2 <211> 2607 <212> PRT <213> Schizochytrium species 4 201038734 <400> 2

Met Asp Thr Arg lie Ala lie Val Gly Met Ser Ala lie Leu Pro Ser 15 10 15Met Asp Thr Arg lie Ala lie Val Gly Met Ser Ala lie Leu Pro Ser 15 10 15

Gly Glu Asn Val Arg Glu Ser Trp Glu Ala lie Arg Asp Gly Leu Asp 20 25 30Gly Glu Asn Val Arg Glu Ser Trp Glu Ala lie Arg Asp Gly Leu Asp 20 25 30

Cys Leu Ser Asp Leu Pro Ala Asp Arg Val Asp Val Thr Ala Tyr Tyr 35 40 45Cys Leu Ser Asp Leu Pro Ala Asp Arg Val Asp Val Thr Ala Tyr Tyr 35 40 45

Asn Pro Glu Lys Thr Thr Lys Asp Lys lie Tyr Cys Lys Arg Gly Gly 50 55 60Asn Pro Glu Lys Thr Thr Lys Asp Lys lie Tyr Cys Lys Arg Gly Gly 50 55 60

Phe lie Pro Glu Tyr Asp Phe Asp Ala Arg Glu Phe Gly Leu Asn Met 65 70 75 80 oPhe lie Pro Glu Tyr Asp Phe Asp Ala Arg Glu Phe Gly Leu Asn Met 65 70 75 80 o

Phe Gin Met Glu Asp Ser Asp Ala Asn Gin Thr lie Ser Leu Leu Lys 85 90 95Phe Gin Met Glu Asp Ser Asp Ala Asn Gin Thr lie Ser Leu Leu Lys 85 90 95

Val Lys Glu Ala Leu Thr Asp Ala Asn lie Pro Ala Phe Ser Ser Gly 100 105 110Val Lys Glu Ala Leu Thr Asp Ala Asn lie Pro Ala Phe Ser Ser Gly 100 105 110

Lys Lys Asn lie Gly Cys Val Leu Gly lie Gly Gly Gly Gin Lys Ala 115 120 125Lys Lys Asn lie Gly Cys Val Leu Gly lie Gly Gly Gly Gin Lys Ala 115 120 125

Ser His Glu Phe Tyr Ser Arg Leu Asn Tyr Val Val Val Asp Lys Val 130 135 140Ser His Glu Phe Tyr Ser Arg Leu Asn Tyr Val Val Val Asp Lys Val 130 135 140

Leu Arg Lys Met Gly Leu Pro Glu Glu Asp Val Ala Ala Ala Val Asp 145 150 155 160Leu Arg Lys Met Gly Leu Pro Glu Glu Asp Val Ala Ala Ala Val Asp 145 150 155 160

Lys Tyr Lys Ala Ser Phe Pro Glu Trp Arg Leu Asp Ser Phe Pro Gly 165 170 175Lys Tyr Lys Ala Ser Phe Pro Glu Trp Arg Leu Asp Ser Phe Pro Gly 165 170 175

Phe Leu Gly Asn Val Thr Ala Gly Arg Cys Cys Asn Thr Phe Asn Met 180 185 190Phe Leu Gly Asn Val Thr Ala Gly Arg Cys Cys Asn Thr Phe Asn Met 180 185 190

Glu Gly Met Asn Cys Val Val Asp Ala Ala Cys Ala Ser Ser Leu lie 195 200 205Glu Gly Met Asn Cys Val Val Asp Ala Ala Cys Ala Ser Ser Leu lie 195 200 205

Ala Val Lys Val Ala lie Glu Glu Leu Leu Tyr Gly Asp Cys Asp Ala 210 215 220Ala Val Lys Val Ala lie Glu Glu Leu Leu Tyr Gly Asp Cys Asp Ala 210 215 220

Met 工le Ala Gly Ala Thr Cys Thr Asp Asn Ser lie Gly Met Tyr Met 225 230 235 240Met Le Ala Gly Ala Thr Cys Thr Asp Asn Ser lie Gly Met Tyr Met 225 230 235 240

Ala Phe Ser Lys Thr Pro Val Phe Ser Thr Asp Pro Ser Val Lys Ala 245 250 255Ala Phe Ser Lys Thr Pro Val Phe Ser Thr Asp Pro Ser Val Lys Ala 245 250 255

Tyr Asp Ala Ala Thr Lys Gly Met Leu lie Gly Glu Gly Ser Ala Met 260 265 270Tyr Asp Ala Ala Thr Lys Gly Met Leu lie Gly Glu Gly Ser Ala Met 260 265 270

Leu Val Leu Lys Arg Tyr Ala Asp Ala Val Arg Asp Gly Asp Thr Val 275 280 285 5 201038734Leu Val Leu Lys Arg Tyr Ala Asp Ala Val Arg Asp Gly Asp Thr Val 275 280 285 5 201038734

His Ala Val lie Lys Gly Cys Ala Ser Ser Ser Asp Gly Lys Ala Ala 290 295 300 e -—i I r o h -—_ T 3 〇 r p r h T r Ty e _—I I y5 1 o G 3His Ala Val lie Lys Gly Cys Ala Ser Ser Ser Asp Gly Lys Ala Ala 290 295 300 e -—I I r o h -—_ T 3 〇 r p r h T r Ty e _—I I y5 1 o G 3

Ser Gly Gin Glu Glu Ala Leu Arg Arg 315 320Ser Gly Gin Glu Glu Ala Leu Arg Arg 315 320

Ala Tyr Ala Arg Ala Asn Val Asp Pro Ala Thr Val Thr Leu Val Glu 325 330 335 Gly His Gly Thr Gly Thr Pro Val Gly Asp Lys lie Glu Leu Thr Ala 340 345 350 Leu Ser Asn Leu Phe Ser Lys Ala Phe Ser Ala Asn Gly Gly Gly Ala 355 360 365 Glu Glu Ala Glu Gin Val Ala Val Gly Ser lie Lys Ser Gin lie Gly 370 375 380Ala Tyr Ala Arg Ala Asn Val Asp Pro Ala Thr Val Thr Leu Val Glu 325 330 335 Gly His Gly Thr Gly Thr Pro Val Gly Asp Lys lie Glu Leu Thr Ala 340 345 350 Leu Ser Asn Leu Phe Ser Lys Ala Phe Ser Ala Asn Gly Gly Gly Ala 355 360 365 Glu Glu Ala Glu Gin Val Ala Val Gly Ser lie Lys Ser Gin lie Gly 370 375 380

His Leu Lys Ala Val Ala Gly Leu Ala Gly Leu Val Lys Val Val Leu 385 390 395 400 cHis Leu Lys Ala Val Ala Gly Leu Ala Gly Leu Val Lys Val Val Leu 385 390 395 400 c

Ala Leu Lys His Lys Thr Leu Pro Gin Thr lie Asn Val Asp Lys Pro 405 410 415 Pro Ser Leu Val Asp Gly Thr Pro lie Gin Gin Ser Pro Leu Tyr Val 420 425 430 Asn Thr Met Asn Arg Pro Trp Phe Thr Pro Val Gly Val Pro Arg Arg 435 440 445 Ala Gly Val Ser Ser Phe Gly Phe Gly Gly Ala Asn Tyr His Ala Val 450 455 460 Leu Glu Glu Phe Glu Pro Glu His Glu Ser Ala Tyr Arg Tyr Asn Asn 465 470 475 480 Leu Pro Gin Val Ala Leu Leu His Ala Gly Asp Val Ala Thr Leu Ala 485 490 495 Ala Thr Val Arg Ala Lys Leu Ala Leu Ala Thr Ala Glu Gin Glu Glu 500 505 510 Ala Arg Val Val Lys Asn Ala Asp Tyr lie Ala Tyr His Arg Phe Leu 515 520 525 Asp Glu Cys Lys Leu Arg Gly Ala Val Pro Gin Ala His Ala Arg Val 530 535 540Ala Leu Lys His Lys Thr Leu Pro Gin Thr lie Asn Val Asp Lys Pro 405 410 415 Pro Ser Leu Val Asp Gly Thr Pro lie Gin Gin Ser Pro Leu Tyr Val 420 425 430 Asn Thr Met Asn Arg Pro Trp Phe Thr Pro Val Gly Val Pro Arg Arg 435 440 445 Ala Gly Val Ser Ser Phe Gly Phe Gly Gly Ala Asn Tyr His Ala Val 450 455 460 Leu Glu Glu Phe Glu Pro Glu His Glu Ser Ala Tyr Arg Tyr Asn Asn 465 470 475 480 Leu Pro Gin Val Ala Leu Leu His Ala Gly Asp Val Ala Thr Leu Ala 485 490 495 Ala Thr Val Arg Ala Lys Leu Ala Leu Ala Thr Ala Glu Gin Glu Glu 500 505 510 Ala Arg Val Val Lys Asn Ala Asp Tyr lie Ala Tyr His Arg Phe Leu 515 520 525 Asp Glu Cys Lys Leu Arg Gly Ala Val Pro Gin Ala His Ala Arg Val 530 535 540

Gly Leu Leu Val Arg Asp Leu Ser Ser Leu lie Ala Val Leu Glu Ala 545 550 555 560Gly Leu Leu Val Arg Asp Leu Ser Ser Leu lie Ala Val Leu Glu Ala 545 550 555 560

Ala Ala Ala Lys Leu Ala Gly Glu Glu 565 a V r 5 h 7 T 5 p r T u _1 G r h T a -—I A r o e 7 s 5 a V r e sAla Ala Ala Lys Leu Ala Gly Glu Glu 565 a V r 5 h 7 T 5 p r T u _1 G r h T a -—I A r o e 7 s 5 a V r e s

r o h 8 T 5 a I—I Ar o h 8 T 5 a I—I A

Gly Glu Ala Ala Phe Arg Val Arg Gly Val Ala Thr 585 590 6 201038734Gly Glu Ala Ala Phe Arg Val Arg Gly Val Ala Thr 585 590 6 201038734

Glu Ala Asn Val Ala Ala Leu 595 Phe Ser Gly Gin Gly Ala Gin Tyr Thr 600 605 His Met Phe Ser Asp Val Ala 610 615 Met Asn Trp Pro Pro Phe Arg Glu Ser 620 Val Ala Ala Met Asp Arg Ala 625 630 Gin Arg Glu Arg Phe Gly Arg Pro Ala 635 640 Lys Arg Val Ser Ser Val Leu 645 Tyr Pro Arg Lys Pro Tyr Gly Asp Glu 650 655 Pro Arg Gin Asp His Lys Glu 660 lie Ser Gin Thr Arg Tyr Ser Gin Pro 665 670 Ala Thr Leu Ala Cys Ser Val 675 Gly Ala Phe Asp lie Phe Lys Ala Ala 680 685 Gly Leu Ala Pro Ser Phe Ala 690 695 Ala Gly His Ser Leu Gly Glu Phe Ala 700 Ala Leu Tyr Ala Ala Gly Ser 705 710 Leu Asp Arg Asp Ala Val Phe Asp Leu 715 720 Val Cys Ala Arg Ala Lys Ala - 725 Met Ser Asp Phe Thr Ala Gin Ala Ser 730 735 Ser Ser Gly Gly Ala Met Ala " 740 Ala Val lie Gly Ala Lys Ala Asp Gin 745 750 Leu Ser Leu Gly Gly Ala Pro 755 Asp Val Trp Leu Ala Asn Ser Asn Ser 760 765 Pro Ser Gin Thr Val lie Thr 770 775 Gly Thr Ala Glu Ala Val Ala Ala Ala 780 Ser Asp Lys Leu Arg Cys Ser 785 790 Gly Asn Phe Arg Val Val Pro Leu Ala 795 800 Cys Glu Ala Ala Phe His Ser 805 Pro His Met Arg Gly Ala Glu Gin Thr 810 815 Phe Ala Ser Ala Leu Ala Gin 820 Ala Pro Val Ser Ala Pro Ala Ala Ala 825 830 Arg Phe Tyr Ser Asn Val Thr 835 Gly Gly Ala Ala Val Thr Ser Pro Ala 840 845Glu Ala Asn Val Ala Ala Leu 595 Phe Ser Gly Gin Gly Ala Gin Tyr Thr 600 605 His Met Phe Ser Asp Val Ala 610 615 Met Asn Trp Pro Pro Phe Arg Glu Ser 620 Val Ala Ala Met Asp Arg Ala 625 630 Gin Arg Glu Arg Phe Gly Arg Pro Ala 635 640 Lys Arg Val Ser Ser Val Leu 645 Tyr Pro Arg Lys Pro Tyr Gly Asp Glu 650 655 Pro Arg Gin Asp His Lys Glu 660 lie Ser Gin Thr Arg Tyr Ser Gin Pro 665 670 Ala Thr Leu Ala Cys Ser Val 675 Gly Ala Phe Asp lie Phe Lys Ala Ala 680 685 Gly Leu Ala Pro Ser Phe Ala 690 695 Ala Gly His Ser Leu Gly Glu Phe Ala 700 Ala Leu Tyr Ala Ala Gly Ser 705 710 Leu Asp Arg Asp Ala Val Phe Asp Leu 715 720 Val Cys Ala Arg Ala Lys Ala - 725 Met Ser Asp Phe Thr Ala Gin Ala Ser 730 735 Ser Ser Gly Gly Ala Met Ala " 740 Ala Val lie Gly Ala Lys Ala Asp Gin 745 750 Leu Ser Leu Gly Gly Ala Pro 755 Asp Val Trp Leu Ala Asn Ser Asn Ser 760 765 Pro Ser Gin Thr Val lie Thr 770 775 Gly Thr Ala Glu Ala Val Ala Ala Ala 780 Ser Asp Lys Leu Arg Cys Ser 785 790 Gly Asn Phe Arg Val Val EZ Phe Tyr Ser Asn Val Thr 835 Gly Gly Ala Ala Val Thr Ser Pro Ala 840 845

Asp Val Lys Thr Asn Leu Gly Lys His Met Thr Ser Pro Val Gin Phe 850 855 860Asp Val Lys Thr Asn Leu Gly Lys His Met Thr Ser Pro Val Gin Phe 850 855 860

Val Gin Gin Val Arg Ala Met 865 870 His Ala Ala Gly Ala Arg Val Phe Val 875 880 Glu Phe Gly Pro Lys Gin Val 885 Leu Ser Arg Leu Val Lys Glu Thr Leu 890 895 7 201038734Val Gin Gin Val Arg Ala Met 865 870 His Ala Ala Gly Ala Arg Val Phe Val 875 880 Glu Phe Gly Pro Lys Gin Val 885 Leu Ser Arg Leu Val Lys Glu Thr Leu 890 895 7 201038734

Gly Glu Ala Gly Asp Val Val Thr Val Ala Val Asn Pro Asp Ser Ala 900 905 910Gly Glu Ala Gly Asp Val Val Thr Val Ala Val Asn Pro Asp Ser Ala 900 905 910

Lys Asp Ser Asp Thr Gin Leu Arg Gin Ala Ala Leu Thr Leu Ala Val 915 920 925Lys Asp Ser Asp Thr Gin Leu Arg Gin Ala Ala Leu Thr Leu Ala Val 915 920 925

Ala Gly Val Pro Leu Lys Asp Phe Asp Arg Trp Gin Leu Pro Asp Ala 930 935 940Ala Gly Val Pro Leu Lys Asp Phe Asp Arg Trp Gin Leu Pro Asp Ala 930 935 940

Thr Arg Leu Glu Pro Val Lys Lys Lys Lys Thr Thr Leu Arg Leu Ser 945 950 955 960Thr Arg Leu Glu Pro Val Lys Lys Lys Lys Thr Thr Leu Arg Leu Ser 945 950 955 960

Ala Ala Thr Tyr Val Ser Ala Lys Thr Leu Arg Gin Arg Glu Ala Val 965 970 975Ala Ala Thr Tyr Val Ser Ala Lys Thr Leu Arg Gin Arg Glu Ala Val 965 970 975

Leu Asn Asp Gly Tyr Thr Val Ser Gly Ala Thr Ala Val Val Lys Glu 980 985 990Leu Asn Asp Gly Tyr Thr Val Ser Gly Ala Thr Ala Val Val Lys Glu 980 985 990

Val Asp Thr Ala Asn Glu Glu Arg Leu Val Arg Gin Ala Gin Asp Leu 995 1000 1005Val Asp Thr Ala Asn Glu Glu Arg Leu Val Arg Gin Ala Gin Asp Leu 995 1000 1005

Gin Arg Gin Leu Ala Glu Ala Ser Thr Ala Ala Gin Ala Ala Gin 1010 1015 1020Gin Arg Gin Leu Ala Glu Ala Ser Thr Ala Ala Gin Ala Ala Gin 1010 1015 1020

Ser Lys Val Ala Glu Leu Glu Arg Thr lie Gin Asp Leu Glu Arg 1025 1030 1035Ser Lys Val Ala Glu Leu Glu Arg Thr lie Gin Asp Leu Glu Arg 1025 1030 1035

Lys Val Gin Gin Gin Gin Gin Glu Lys Gly Glu Asn Ser Asp Ser 1040 1045 1050Lys Val Gin Gin Gin Gin Gin Glu Lys Gly Glu Asn Ser Asp Ser 1040 1045 1050

Asn Ala Ala Ala Glu Val Leu Arg Arg His Lys Glu Leu Leu Gin 1055 1060 1065Asn Ala Ala Ala Glu Val Leu Arg Arg His Lys Glu Leu Leu Gin 1055 1060 1065

Arg Met Leu Gin Asp Cys Asp Glu Gin Ala Val Pro Val Ala Thr 1070 1075 1080Arg Met Leu Gin Asp Cys Asp Glu Gin Ala Val Pro Val Ala Thr 1070 1075 1080

Val Val Pro Thr Pro Thr Ser Ser Pro Thr Pro Thr Ser Ser Pro 1085 1090 1095Val Val Pro Thr Pro Thr Ser Ser Pro Pro Ser Pro 1085 1090 1095

Val Ser Gly Asn Ser Lys Ser Thr Arg Gly Ser Ala Asp Leu Gin 1100 1105 1110Val Ser Gly Asn Ser Lys Ser Thr Arg Gly Ser Ala Asp Leu Gin 1100 1105 1110

Ala Leu Leu Ala Lys Ala Glu Thr Val Val Met Ala Val Leu Ala 1115 1120 1125Ala Leu Leu Ala Lys Ala Glu Thr Val Val Met Ala Val Leu Ala 1115 1120 1125

Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala Asp Met Asp 1130 1135 1140Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala Asp Met Asp 1130 1135 1140

Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 1145 1150 1155Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 1145 1150 1155

Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp Val 1160 1165 1170Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp Val 1160 1165 1170

Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 8 201038734 1175 1180 1185Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 8 201038734 1175 1180 1185

Met Lys Ala Glu lie Val Ala Ala Ser Gly Gly Ser 1190 1195 1200Met Lys Ala Glu lie Val Ala Ala Ser Gly Gly Ser 1190 1195 1200

Val Pro Ser Ala Pro Ala Ala Ser Ala Ala Pro Thr 1205 1210 1215Val Pro Ser Ala Pro Ala Ala Ser Ala Ala Pro Thr 1205 1210 1215

Ser Thr Ala Pro Ser Ala Asp Leu Gin Ala Leu Leu 1220 1225 1230Ser Thr Ala Pro Ser Ala Asp Leu Gin Ala Leu Leu 1220 1225 1230

Glu Thr Val Val Met Ala Val Leu Ala Ala Lys Thr 1235 1240 1245Glu Thr Val Val Met Ala Val Leu Ala Ala Lys Thr 1235 1240 1245

Ala Asp Met Val Glu Ala Asp Met Asp Leu Glu Ala 1250 1255 1260 Ο lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu 1265 1270 1275Ala Asp Met Val Glu Ala Asp Met Asp Leu Glu Ala 1250 1255 1260 Ο lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu 1265 1270 1275

Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu 1280 1285 1290Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu 1280 1285 1290

Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala 1295 1300 1305Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala 1295 1300 1305

Ala Ala Ser Ala Gly Ser Ala Pro Ala Pro Ala Val 1310 1315 1320Ala Ala Ser Ala Gly Ser Ala Pro Ala Pro Ala Val 1310 1315 1320

Pro Ala Ala Ser Ala Ala Pro Thr Pro Ala Ala Ser 1325 1330 1335Pro Ala Ala Ser Ala Ala Pro Thr Pro Ala Ala Ser 1325 1330 1335

Ser Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu 1340 1345 1350Ser Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu 1340 1345 1350

Met Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala 1355 1360 1365Met Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala 1355 1360 1365

Glu Ala Asp Met Asp Leu Glu Ala Glu Leu Gly lie 1370 1375 1380Glu Ala Asp Met Asp Leu Glu Ala Glu Leu Gly lie 1370 1375 1380

Lys Arg Val Glu lie Leu Ser Glu Val Gin Gly Gin 1385 1390 1395Lys Arg Val Glu lie Leu Ser Glu Val Gin Gly Gin 1385 1390 1395

Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg 1400 1405 1410Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg 1400 1405 1410

Glu Val Val Asp Ala Met Lys Ala Glu lie Val Ala 1415 1420 1425Glu Val Val Asp Ala Met Lys Ala Glu lie Val Ala 1415 1420 1425

Gly Ser Ala Pro Ala Pro Ala Val Pro Ser Ala Pro 1430 1435 1440Gly Ser Ala Pro Ala Pro Ala Val Pro Ser Ala Pro 1430 1435 1440

Ala Ala Pro Thr Pro Ala Ala Ala Thr Ala Pro Ser 1445 1450 1455Ala Ala Pro Thr Pro Ala Ala Ala Thr Ala Pro Ser 1445 1450 1455

Ala Pro Ala Pro Ala Ala Ser Lys Ala Gly Tyr Glu Glu Leu Gly Val Gin Gly Ser Arg Thr Glu He Val Pro Ser Ala Thr Ala Pro Thr Val Val Asp Met Val Asp Ser lie Leu Gly Val Thr Val Gly Ala Ser Gly Ala Ala Ser Ala Asp Leu 9 201038734Ala Pro Ala Pro Ala Ala Ser Lys Ala Gly Tyr Glu Glu Leu Gly Val Gin Gly Ser Arg Thr Glu He Val Pro Ser Ala Thr Ala Pro Thr Val Val Asp Met Val Asp Ser lie Leu Gly Val Thr Val Gly Ala Ser Gly Ala Ala Ser Ala Asp Leu 9 201038734

Gin Ala 1460Gin Ala 1460

Leu Leu Ala Lys Ala Glu Thr Val Val Met Ala Val Leu 1465 1470Leu Leu Ala Lys Ala Glu Thr Val Val Met Ala Val Leu 1465 1470

Ala Ala 1475Ala Ala 1475

Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala Asp Met 1480 1485Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala Asp Met 1480 1485

Asp Leu 1490Asp Leu 1490

Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu 1495 1500 lie Leu 1505Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu 1495 1500 lie Leu 1505

Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 1510 1515Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 1510 1515

Val Asp 1520Val Asp 1520

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp 1525 1530Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp 1525 1530

Ala Met 1535Ala Met 1535

Lys Ala Glu lie Val Ala Ala Ser Ala Gly Ser Ala Pro 1540 1545Lys Ala Glu lie Val Ala Ala Ser Ala Gly Ser Ala Pro 1540 1545

Ala Pro 1550Ala Pro 1550

Ala Val Pro Ser Ala Pro Ala Ala Ser Ala Ala Pro Thr 1555 1560Ala Val Pro Ser Ala Pro Ala Ala Ser Ala Ala Pro Thr 1555 1560

Pro Ala 1565Pro Ala 1565

Ala Ser Thr Ala Pro Ser Ala Asp Leu Gin Ala Leu Leu 1570 1575Ala Ser Thr Ala Pro Ser Ala Asp Leu Gin Ala Leu Leu 1570 1575

Ser Lys 1580Ser Lys 1580

Ala Glu Thr Val Val Met Ala Val Leu Ala Ala Lys Thr 1585 1590Ala Glu Thr Val Val Met Ala Val Leu Ala Ala Lys Thr 1585 1590

Gly Tyr 1595Gly Tyr 1595

Glu Ala Asp Met Val Glu Ala Asp Met Asp Leu Glu Ala 1600 1605Glu Ala Asp Met Val Glu Ala Asp Met Asp Leu Glu Ala 1600 1605

Glu Leu 1610Glu Leu 1610

Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu 1615 1620Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu 1615 1620

Val Gin 1625Val Gin 1625

Gly Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu 1630 1635Gly Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu 1630 1635

Ser Arg 1640Ser Arg 1640

Thr Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala 1645 1650Thr Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala 1645 1650

Glu lie 1655Glu lie 1655

Val Ala Ala Ser Gly Gly Ser Ala Pro Ala Ala Ala Val 1660 1665Val Ala Ala Ser Gly Gly Ser Ala Pro Ala Ala Ala Val 1660 1665

Pro Ser 1670Pro Ser 1670

Ala Pro Ala Ala Ser Ala Ala Pro Thr Pro Ala Thr Ala 1675 1680Ala Pro Ala Ala Ser Ala Ala Pro Thr Pro Ala Thr Ala 1675 1680

Pro Ser 1685Pro Ser 1685

Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu Thr Val 1690 1695Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu Thr Val 1690 1695

Val Met 1700Val Met 1700

Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met 1705 1710Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met 1705 1710

Val Glu 1715Val Glu 1715

Ala Asp Met Asp Leu Glu Ala Glu Leu Gly 工le Asp Ser 1720 1725 lie Lys 1730Ala Asp Met Asp Leu Glu Ala Glu Leu Gly gong le Asp Ser 1720 1725 lie Lys 1730

Arg Val Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly 1735 1740 10 201038734 ΟArg Val Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly 1735 1740 10 201038734 Ο

Val Glu 1745 Ala Lys Asp Val Asp 1750 Ala Leu Ser Arg Thr 1755 Arg Thr Val Gly Glu 1760 Val Val Asp Ala Met 1765 Lys Ala Glu lie Val 177 0 Ala Ala Ser Gly Gly 1775 Ser Ala Pro Ala Ala 1780 Pro Ser Ala Pro Ala 1785 Leu Leu Pro Thr Leu 1790 Phe Gly Ser Glu Cys 1795 Glu Asp Leu Ser Leu 1800 Thr Phe Pro Val He 1805 Thr Thr Leu Pro Leu 1810 Pro Ala Glu Leu Val 1815 Leu Ala Glu Gly Gly 1820 Ala Arg Pro Val Val 1825 Val Val Asp Asp Gly 1830 Ser Ala Leu Thr Ser 1835 Ser Leu Val Ser Ser 1840 Leu Gly Asp Arg Ala 1845 Val Leu Leu Gin Val 1850 Gin Ser Ser Ser Ala 1855 Cys Ser Pro Arg Ser 1860 Thr Thr His Lys Leu 1865 Val Thr Val Ala Asp 1870 Arg Ser Glu Ala Ala 1875 Leu Gin Ala Ala Leu 1880 Thr Ser Val Glu Ala 1885 Gin Phe Gly Lys Val 1890 Gly Gly Phe Val Phe 1895 Gin Phe Gly Asp Asp 1900 Asp Val Gin Ala Gin 1905 Leu Gly Trp Ala Leu 1910 Leu Ala Ala Lys His 1915 Leu Lys Thr Ser Leu 1920 Ser Glu Gin lie Glu 1925 Gly Gly Arg Thr Phe 1930 Phe Val Ala Val Ala 1935 Arg Leu Asp Gly Gin 1940 Leu Gly Leu Ser Gly 1945 Lys Ser Thr Thr Ala 1950 Thr Val Asp Leu Ser 1955 Arg Ala Gin Gin Gly 1960 Ser Val Phe Gly Leu 1965 Cys Lys Thr Leu Asp 1970 Leu Glu Trp Pro Ala 1975 Val Phe Cys Arg Gly 1980 lie Asp Leu Ala Ala 1985 Asp Leu Asp Ala Ala 1990 Gin Ala Ala Arg Cys 1995 Leu Leu Gly Glu Leu 2000 Ser Asp Pro Asp Val 2005 Ala Val Arg Glu Ser 2010 Gly Tyr Ser Ala Ser 2015 Gly Gin Arg Cys Thr 2020 Thr Thr Thr Lys Ser 2025 Leu Thr Thr 11 201038734Val Glu 1745 Ala Lys Asp Val Asp 1750 Ala Leu Ser Arg Thr 1755 Arg Thr Val Gly Glu 1760 Val Val Asp Ala Met 1765 Lys Ala Glu lie Val 177 0 Ala Ala Ser Gly Gly 1775 Ser Ala Pro Ala Ala 1780 Pro Ser Ala Pro Ala 1785 Leu Leu Pro Thr Leu 1790 Phe Gly Ser Glu Cys 1795 Glu Asp Leu Ser Leu 1800 Thr Phe Pro Val He 1805 Thr Thr Leu Pro Leu 1810 Pro Ala Glu Leu Val 1815 Leu Ala Glu Gly Gly 1820 Ala Arg Pro Val Val 1825 Val Val Asp Asp Gly 1830 Ser Ala Leu Thr Ser 1835 Ser Leu Val Ser Ser 1840 Leu Gly Asp Arg Ala 1845 Val Leu Leu Gin Val 1850 Gin Ser Ser Ser Ala 1855 Cys Ser Pro Arg Ser 1860 Thr Thr His Lys Leu 1865 Val Thr Val Ala Asp 1870 Arg Ser Glu Ala Ala 1875 Leu Gin Ala Ala Leu 1880 Thr Ser Val Glu Ala 1885 Gin Phe Gly Lys Val 1890 Gly Gly Phe Val Phe 1895 Gin Phe Gly Asp Asp 1900 Asp Val Gin Ala Gin 1905 Leu Gly Trp Ala Leu 1910 Leu Ala Ala Lys His 1915 Leu Lys Thr Ser Leu 1920 Ser Glu Gin lie Glu 1925 Gly Gly Arg Thr Phe 1930 Phe Val Ala Val Ala 1935 Arg Leu Asp Gly Gin 1940 Leu Gly Leu Ser Gly 1945 Lys Ser Thr Thr Ala 1950 Thr Val Asp Leu Ser 1955 Arg Ala Gin Gin Gly 1960 Ser Val Phe Gly Leu 1965 Cys Lys Thr Leu Asp 1970 Leu Glu Trp Pro Ala 1975 Val Phe Cys Arg Gly 1980 lie Asp Leu Ala Ala 1985 Asp Leu Asp Ala Ala 1990 Gin Ala Ala Arg Cys 1995 Leu Leu Gly Glu Leu 2000 Ser Asp Pro Asp Val 2005 Ala Val Arg Glu Ser 2010 Gly Tyr Ser Ala Ser 2015 Gly Gin Arg Cys Thr 2020 Thr Thr Thr Lys Ser 2025 Leu Thr Thr 11 201038734

Gly Lys Pro His Gin Pro lie Ser Ser Ser Asp Leu Phe Leu Val 2030 2035 2040Gly Lys Pro His Gin Pro lie Ser Ser Ser Asp Leu Phe Leu Val 2030 2035 2040

Ser Gly Gly Ala Arg Gly lie Thr Pro Leu Cys Val Arg Glu Leu 2045 2050 2055Ser Gly Gly Ala Arg Gly lie Thr Pro Leu Cys Val Arg Glu Leu 2045 2050 2055

Ala Gin Arg Val Gly Gly Gly Thr Tyr Val Leu lie Gly Arg Ser 2060 2065 2070Ala Gin Arg Val Gly Gly Gly Thr Tyr Val Leu lie Gly Arg Ser 2060 2065 2070

Glu Leu Pro Thr Thr Glu Pro Ala Trp Ala Val Gly Val Glu Ser 2075 2080 2085Glu Leu Pro Thr Thr Glu Pro Ala Trp Ala Val Gly Val Glu Ser 2075 2080 2085

Gly Lys Pro Leu Glu Lys Ala Ala Leu Ala Phe Leu Lys Ala Glu 2090 2095 2100Gly Lys Pro Leu Glu Lys Ala Ala Leu Ala Phe Leu Lys Ala Glu 2090 2095 2100

Phe Ala Ala Gly Arg Gly Ala Lys Pro Thr Pro Met Leu His Lys 2105 2110 2115Phe Ala Ala Gly Arg Gly Ala Lys Pro Thr Pro Met Leu His Lys 2105 2110 2115

Lys Leu Val Gly Ala Val Val Gly Ala Arg Glu Val Arg Ala Ser 2120 2125 2130Lys Leu Val Gly Ala Val Val Gly Ala Arg Glu Val Arg Ala Ser 2120 2125 2130

Leu Ala Glu lie Thr Ala Gin Gly Ala Thr Ala Val Tyr Glu Ser 2135 2140 2145Leu Ala Glu lie Thr Ala Gin Gly Ala Thr Ala Val Tyr Glu Ser 2135 2140 2145

Cys Asp Val Ser Ser Ala Ala Lys Val Arg Glu Met Val Glu Arg 2150 2155 2160Cys Asp Val Ser Ser Ala Ala Lys Val Arg Glu Met Val Glu Arg 2150 2155 2160

Val Gin Gin Gin Gly Gly Arg Arg Val Ser Gly Val Phe His Ala 2165 2170 2175Val Gin Gin Gin Gly Gly Arg Arg Val Ser Gly Val Phe His Ala 2165 2170 2175

Ser Gly Val Leu Arg Asp Lys Leu Val Glu Asn Lys Ser Leu Ala 2180 2185 2190Ser Gly Val Leu Arg Asp Lys Leu Val Glu Asn Lys Ser Leu Ala 2180 2185 2190

Asp Phe Ser Ala Val Tyr Asp Thr Lys Val Gly Gly Leu lie Asn 2195 2200 2205Asp Phe Ser Ala Val Tyr Asp Thr Lys Val Gly Gly Leu lie Asn 2195 2200 2205

Leu Leu Ala Cys Val Asp Leu Ala Gin Leu Arg His Leu Val Leu 2210 2215 2220Leu Leu Ala Cys Val Asp Leu Ala Gin Leu Arg His Leu Val Leu 2210 2215 2220

Phe Ser Ser Leu Ala Gly Phe His Gly Asn Val Gly Gin Ser Asp 2225 2230 2235Phe Ser Ser Leu Ala Gly Phe His Gly Asn Val Gly Gin Ser Asp 2225 2230 2235

Tyr Ala Met Ala Asn Glu Ala Leu Asn Lys Leu Ala Ala His Leu 2240 2245 2250Tyr Ala Met Ala Asn Glu Ala Leu Asn Lys Leu Ala Ala His Leu 2240 2245 2250

Ser Ala Val His Pro Gin Leu Cys Ala Arg Ser lie Cys Phe Gly 2255 2260 2265Ser Ala Val His Pro Gin Leu Cys Ala Arg Ser lie Cys Phe Gly 2255 2260 2265

Pro Trp Asp Gly Gly Met Val Thr Pro Ala Leu Lys Ala Asn Phe 2270 2275 2280 lie Arg Met Gly lie Gin lie lie Pro Arg Gin Gly Gly Ala Gin 2285 2290 2295Pro Trp Asp Gly Gly Met Val Thr Pro Ala Leu Lys Ala Asn Phe 2270 2275 2280 lie Arg Met Gly lie Gin lie lie Pro Arg Gin Gly Gly Ala Gin 2285 2290 2295

Thr Val Ala Asn Met Leu Val Ser Ser Ser Pro Gly Gin Leu Leu 12 201038734 2300 2305 2310Thr Val Ala Asn Met Leu Val Ser Ser Ser Pro Gly Gin Leu Leu 12 201038734 2300 2305 2310

Val Gly Asn Trp Gly Val Pro 2315 2320Val Gly Asn Trp Gly Val Pro 2315 2320

Pro Val Val Pro Ser 2325Pro Val Val Pro Ser 2325

His Thr Val Leu Gin Thr Leu 2330 2335His Thr Val Leu Gin Thr Leu 2330 2335

Arg Gin Ser Asp Asn 2340Arg Gin Ser Asp Asn 2340

Asp Ser His Val lie Gin Gly 2345 2350Asp Ser His Val lie Gin Gly 2345 2350

Arg Arg Val Leu Pro 2355Arg Arg Val Leu Pro 2355

Ala Val Gly Tyr Met Ala His 2360 2365Ala Val Gly Tyr Met Ala His 2360 2365

Gin Ala Gin Ser lie 2370Gin Ala Gin Ser lie 2370

His Gin Leu Trp Ala Val Glu 2375 2380His Gin Leu Trp Ala Val Glu 2375 2380

Asp Ala Gin Leu Phe 2385Asp Ala Gin Leu Phe 2385

Ala lie Asp Asn Gly Ala Asp 2390 2395Ala lie Asp Asn Gly Ala Asp 2390 2395

Val Pro Val Arg Val 2400Val Pro Val Arg Val 2400

Arg Arg 2405Arg Arg 2405

Lys Glu Glu Gin Glu 2410Lys Glu Glu Gin Glu 2410

Asp Ala Gly Lys Val 2415Asp Ala Gly Lys Val 2415

Val Gin 2420Val Gin 2420

Val Leu Leu Lys Ser 2425Val Leu Leu Lys Ser 2425

Gin Val Asn Gly Lys 2430Gin Val Asn Gly Lys 2430

Ala Tyr 2435Ala Tyr 2435

Lys Ala Thr Val Val 2440Lys Ala Thr Val Val 2440

Leu Ser Pro Ala Pro 2445Leu Ser Pro Ala Pro 2445

Val lie 2450Val lie 2450

Thr Arg Asp Phe Asp 2455Thr Arg Asp Phe Asp 2455

Leu Thr Pro Asp Pro 2460Leu Thr Pro Asp Pro 2460

Glu His 2465Glu His 2465

Asp Leu Tyr Asp Gly 2470Asp Leu Tyr Asp Gly 2470

Lys Thr Leu Phe His 2475Lys Thr Leu Phe His 2475

Phe Gin Gly lie Glu Gin Val 2480 2485Phe Gin Gly lie Glu Gin Val 2480 2485

Leu Ser Ala Thr Pro 2490Leu Ser Ala Thr Pro 2490

Thr Ala Lys Cys Arg Asn Leu 2495 2500Thr Ala Lys Cys Arg Asn Leu 2495 2500

Pro Leu Thr Pro Glu 2505Pro Leu Thr Pro Glu 2505

Gin Phe Val Val Asn Leu Ser 2510 2515Gin Phe Val Val Asn Leu Ser 2510 2515

Gin Gin Asp Pro Phe 2520 lie Ala Phe Gin Ala Met Leu 2525 2530Gin Gin Asp Pro Phe 2520 lie Ala Phe Gin Ala Met Leu 2525 2530

Val Trp Ala Arg Met 2535Val Trp Ala Arg Met 2535

Ser Ala Ala Leu Pro Asn Asn 2540 2545Ser Ala Ala Leu Pro Asn Asn 2540 2545

Cys Glu Arg Phe Asp 2550Cys Glu Arg Phe Asp 2550

Pro Met Ala Pro Gly Ala Thr 2555 2560Pro Met Ala Pro Gly Ala Thr 2555 2560

Tyr Tyr Thr Ser Val 2565Tyr Tyr Thr Ser Val 2565

Ser Ala Ser Pro Leu Val Asp 2570 2575Ser Ala Ser Pro Leu Val Asp 2570 2575

Ser Val Cys Lys Cys 2580Ser Val Cys Lys Cys 2580

Ala Thr Glu Pro Phe Leu Met Thr Leu Tyr Ala Gly Lys Gly lie Glu Leu Ser Lys Val Lys Ser Val Pro Arg Pro Ser Ala Cys Thr Gly Lys Ala Lys Gin Leu Gin Arg Gly Gin Ala Asp Leu Arg Gin Phe Tyr Lys Lys Leu Ala Thr Val Ala 13 201038734Ala Thr Glu Pro Phe Leu Met Thr Leu Tyr Ala Gly Lys Gly lie Glu Leu Ser Lys Val Lys Ser Val Pro Arg Pro Ser Ala Cys Thr Gly Lys Ala Lys Gin Leu Gin Arg Gly Gin Ala Asp Leu Arg Gin Phe Tyr Lys Lys Leu Ala Thr Val Ala 13 201038734

Met His 2585 Asp Glu Gin Gly Glu 2590 Val Tyr Phe Ser Ala Arg Ala Ser 2595 Val Val 2600 Leu Asn Lys Thr Leu 2605 Thr Tyr <210〉 3 <211> 5778 <212> DNA <213> 裂殖壺菌物種 <400〉 3 atgccgtgcg ataacattgc ggtcgtgggc atggcggtgc caggacgagt tctgggatac gctgatgcgt aaggagatca gagcgcctcg gtacgcgcta ccgcgacctc cacttccacc gacaccttct gcaacgatcg ctacggctgc gtcgatgcca ctcctcgccg acctggcccg gcgcgccctg ctcgacgccg agcaccaccg ccaacctacg cgacttcggc atcgtgagcg gacaatctgc agggcgagct gctcaatctg taccaagtgc gcccagcgct tccgcgactc gcgcccctgg tcggagcgcc gccagcgacc cgcgcgtgta ctccgacccg gcgtccttcg gggcccgtgc gctacagcct cgatgcagcc tgcgcgtcgg gcgtccgacc acttgctctc gcgcagcgcg gacgtgatgc ccggacccgt tcttcattct ctcggggttc tccaccttcc ccggacgata acccactgtc cgtgccgctg cggcagggca gagggcggcg ccatcatggt gctgaagcgc ctcgaggacg atctacggca ccttgctcgg cacgagtctg agcaacgccg ccgcacctgc cgagcgagaa gtcgtgcatg gaggacctgt ccaagcgagg tgcagtacgt ggagtgccac gccacgggca gaggtagagg cgctgcgcca ctgctttcga ggtaacacgg tccaccaagg gcaactttgg ccacactctc gtggcggccg gtgctgctgt cgatgcagca cggcacgatc ccgcccacgc tgcatcgacc cgctcgtcgt ggacgaggcc atcccttggc cgggcaggca aaccaggcga tgagctcaag tgcgcctcgc ggaaccaacg cgcactgtgt cttccgtgag caccgccaaa tcgccggtgc ttcccgaggt gactcctgga ccgattgcca tttggtaccc tcaagggcct ggacgcgttt gagcaggcca gccagcgacc tgccgagcaa gcgctggcgg ttcctgggcg gccatgggcc tcgacgccgt gccgcgcggg tgctacgtgc aagcggctgc ggtcgccgat gatccctgag gacgtcctgc gtggctacga tggaccgcgc gctgcaggac gctggaatgg gtgctggtgg ggctcggcac ggacaccgag ctgtaccggc agtatgccgg atgcaagaac 60 actcgagccc gatctcggcg 120 cgcagcgcag caagtacgcc 180 gcgtcgacaa cgagcacgac 240 gaattaacct cgacgacgcc 300 gctgcctgtc gttccccatg 360 atgtggagaa ccgcgtgggc 420 cgcgcgctgt ctcgcccgag 480 tggccaacca gctcggcctg 540 cgctgtactg cctcaagctg 600 tgtgcggcgc cacatgcttt 660 aggcgatgcc gctgggcgga 720 gccagggcct gacgcccgga 780 ccgtgcgcga cggcgaccgc 840 ggtgcggcct gccgctgagc 900 acacgagcgt cggcatcgac 960 ctccgcaggg cgacgtcgtg 1020 accacccgcc gcgcatgggc 1080 ggttcgcagg catggccaag 1140 ccggtgtcga ccgctccaac 1200 cgtactcgtc ggcgcaggcg 1260 tctccgcctt tggctttggt 1320 ttgctgctac tgcgacagcc 1380 tcatcgggat ggacgcgacg 1440 tctacaaggg cacggacggc 1500 ccgacacgga cttcttgacc 1560 gcgacgtgga cgtggactac 1620 gcccgcaaca gctgctggcg 1680 cgacgggagg caaggtggcg 17 40 accgcgcgcg cgtgacactc 1800Met His 2585 Asp Glu Gin Gly Glu 2590 Val Tyr Phe Ser Ala Arg Ala Ser 2595 Val Val 2600 Leu Asn Lys Thr Leu 2605 Thr Tyr <210> 3 <211> 5778 <212> DNA <213> Thraustochytrium species < 400> 3 atgccgtgcg ataacattgc ggtcgtgggc atggcggtgc caggacgagt tctgggatac gctgatgcgt aaggagatca gagcgcctcg gtacgcgcta ccgcgacctc cacttccacc gacaccttct gcaacgatcg ctacggctgc gtcgatgcca ctcctcgccg acctggcccg gcgcgccctg ctcgacgccg agcaccaccg ccaacctacg cgacttcggc atcgtgagcg gacaatctgc agggcgagct gctcaatctg taccaagtgc gcccagcgct tccgcgactc gcgcccctgg tcggagcgcc gccagcgacc cgcgcgtgta ctccgacccg gcgtccttcg gggcccgtgc gctacagcct cgatgcagcc tgcgcgtcgg gcgtccgacc acttgctctc gcgcagcgcg gacgtgatgc ccggacccgt tcttcattct ctcggggttc tccaccttcc ccggacgata acccactgtc cgtgccgctg cggcagggca gagggcggcg ccatcatggt gctgaagcgc ctcgaggacg atctacggca ccttgctcgg cacgagtctg agcaacgccg ccgcacctgc cgagcgagaa gtcgtgcatg gaggacctgt ccaagcgagg tgcagtacgt ggagtgccac gccacgggca gaggtagagg cgctgcgcca ctgctttcga ggtaaca cgg tccaccaagg gcaactttgg ccacactctc gtggcggccg gtgctgctgt cgatgcagca cggcacgatc ccgcccacgc tgcatcgacc cgctcgtcgt ggacgaggcc atcccttggc cgggcaggca aaccaggcga tgagctcaag tgcgcctcgc ggaaccaacg cgcactgtgt cttccgtgag caccgccaaa tcgccggtgc ttcccgaggt gactcctgga ccgattgcca tttggtaccc tcaagggcct ggacgcgttt gagcaggcca gccagcgacc tgccgagcaa gcgctggcgg ttcctgggcg gccatgggcc tcgacgccgt gccgcgcggg tgctacgtgc aagcggctgc ggtcgccgat gatccctgag gacgtcctgc gtggctacga tggaccgcgc gctgcaggac gctggaatgg gtgctggtgg ggctcggcac ggacaccgag ctgtaccggc agtatgccgg atgcaagaac 60 actcgagccc gatctcggcg 120 cgcagcgcag caagtacgcc 180 gcgtcgacaa cgagcacgac 240 gaattaacct cgacgacgcc 300 gctgcctgtc gttccccatg 360 atgtggagaa ccgcgtgggc 420 cgcgcgctgt ctcgcccgag 480 tggccaacca gctcggcctg 540 cgctgtactg cctcaagctg 600 tgtgcggcgc cacatgcttt 660 aggcgatgcc gctgggcgga 720 gccagggcct gacgcccgga 780 ccgtgcgcga cggcgaccgc 840 ggtgcggcct gccgctgagc 900 acacgagcgt cggcatcgac 960 ctccgcaggg cgacgtcgtg 1020 Accacccgc c gcgcatgggc 1080 ggttcgcagg catggccaag 1140 ccggtgtcga ccgctccaac 1200 cgtactcgtc ggcgcaggcg 1260 tctccgcctt tggctttggt 1320 ttgctgctac tgcgacagcc 1380 tcatcgggat ggacgcgacg 1440 tctacaaggg cacggacggc 1500 ccgacacgga cttcttgacc 1560 gcgacgtgga cgtggactac 1620 gcccgcaaca gctgctggcg 1680 cgacgggagg caaggtggcg 17 40 accgcgcgcg cgtgacactc 1800

14 201038734 Ο ❹ aaggagcggc atcaacgact cgcgtgtcct tcggtctacc gtcgtggtgg cgctccgcca tactttgccg gacgacaagg gtgtcgccgc gtggagctcg gccgagtcgc gccatcggca ctcatcaaga cggccggtgg aagaacccgg tttggcgtgc gatgacgccg aaggtcaacg gacgacccag ggggagacgg ccggagaagc gccgggcgca gaccgcgtcg ctgcaccgcc gagaacggcg cagacggcct tgcctcaccg agcctcggcg cagctcacgc caggccttgc ggctacttgg ttcgtgcgcc gagtgtctgc ggcatgattg gagattttgg ctgcgcgggg gccgactttc gtggggccga tcgacccggc gcggcacctc cgcagtgggg gctgcctcga ccggcgtcga tcagccgaca gcgagggcag tctacgccag tactactcca ccgccgactc agctggagca gcgtgcgcgc cggcgctgtg cgccggtctc gcgagtcgcg tcctcacaga cccccaagct ccgagctggc ctgctgctgt tggctagtca tggagaagga actggatgtc cgttcatgta tgtggccggc actcgtggct ttgacgcgga actacgcgcg agatctccat agcgcctacg gcaagctatg ttcgcgccag tgctgatcgt gcgtgctgga ggcactgccc agattccgga gcagcaacag cgggcatcgt acaacatgcg cgcgttctcg gacgtcgtac ctttacgggc gctgggcaag cctctgtgcc ggaccaccct cggcgccctg tgtcgcgggc agtccacaac gggtcgccat agtgtccaag caacgtggga cctccacaac cgaggcgctg actggcggct cgagtacgcc catcgcgatc gctcctccga cgctttcact cggtgccacg gttggagctg gccatcgggc cggcgagggc tttgcacgag catgccgcgc ccagatcgag cgacgtgctc gctctttgcg cacctcgccg gaacgtgccg ccgcgccgaa caacgactcg gcgcctgggc cgaagtggcg cagccccgtc cagcatcacc cgacaaggtc ctccgccgcg cccgagcagg acgtcgtaca ccgtccttca ttcctgctcg accgccgaga cgcgccaact gtcctcaagc ctcacgtgcg gacgacaacg gcgccgcact ttgctcgcgc gacgtcgggt cgctacctcc tttacttgcc gtcgccagtg actcatgaga cgtggcgaca gcgcacgccg gctcatcgct gcgaccttgt gcagccaagg agcgcctttg cgcagcccct cgcatcaacg gcggtggatg atgttccgca ggggtgcagc ctgtcgcgac gtgtggtcga gcggacgccc atcgagaagg agcagcgcgc gggcggttgc ccctacacgc aagatgtaca gagttcgtgc agccgtgacg gtcagtgaca tgcaggagat tcggcaacct ccgtcaccga acacgcacca acctttacct ttgaggccag gccaggccga ccgcgcagcc agaagagggt tggccaactc accaggtgcc acgcctcggg cggccaaccc cgcgctcgcg cctccgagag gcagcaaccg ccgttgacga aaaccgggtc tgcgcttttt gtttggccct gtgtaccgcg cgccgacacc actacggcgt acaagaccgc ccgactcgca cgggcatctt ccaaggcgtg gcaactgcgg cacagctggc ccgtggagtc cgatcgggcc tgatcgccgg cgccgatgcc cgggcatcgc cctcggtcac agaagttgta gccacgatgt ttcttggcaa gatggactac cgtggccacg aggcgcaaac ggtggacgcc caaggcgcgc cgccgacggg cgttggctca cgctgaagcc ggtggagatg gccgctgagc gggctcggtg cgccgcgagc gcagtgggag tgcctggctg cgggtcctgc cctctcgctg tatcatggcc tgctactgac gcggctcgta gctgacaacg aagcgccaag tgtgaccagc cgggctcgac ggcgctgtgg gcgcgccgtg cgtgtccatc cttcggcctc cctgtcggac ggtggagttc cttctggcag cgacaaccgc caaacctgcc cgtcaagcaa gcacatccac caacgccgag cacgcgcatc cttcgtcgag ggctgccacc 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 15 201038734 ccgcatgtct ccgtggcgct ggaccgcccc agtgagtcgg cgtggacgca gaccctcaag 4140 tcgctggcgc tgctgaccgc ccaccgcgtg cccctgcaca acccgactct gtttgcggac 4200 ctgtaccacc ccacgttcct gacggctatc gactctgcga tgcaggagcc cccgcccaag 4260 cccaaccgct tccttcgcag cgtagaggtc aacgggtact tttgccccga cggcatcagc 4320 aagcaggttg ctgctgcaag tgccaaaccc tcgacgcatt gcatggttcg tttgcaccca 4380 gccaaggcag ttgtggttgc tgctgctggt gctgtggttg ctgattcgac gcccgtggtc 4440 aaggccaagc agacgtcgtc gtcgttgttg gttggggatg acgcctttct gcgctgctac 4500 gacgtggact ggccgctcta catgggcgcc atggcggaag gcatctcgtc ggtagacctg 4560 gtggtcgctg ccgccgaggc ccgcatgctg gcatcattcg gagcggcccg cttgcctatg 4620 gaccaggtgg aactccagat ccgtgagatc cagcaacgca cctccaacgc ctttgctgtc 4680 aacctgatgc cgggtcctga cgaggccgcg acggtggacg cgctgctgcg cacgggcgtc 47 40 tcaatcgtcg aggcatcggg ctacaccggc gcgctctctg cagacctggt gcgctaccgt 4800 gtcacgggtc tgcgacgaac tagttgcggt gcttctgtgt cggcgactca ccgtgtggtc 4860 gccaaggtgt cgcgcaccga ggtggccgag cactttctgc gcccggcgcc ggccgccgta 4920 ctagaggctt tggtcgccgc caaacagatt acgcccgagc aggccgcgct ggccagccgc 4980 gtcgccatgg ccgacgacgt cgcggtggag gccgactcgg gcgggcacac cgacaaccga 5040 ccgatccacg tgctgctgcc gctcgtggtg gcgcagcgca accgctggcg ccacctggtg 5100 gacacgccag tgcgcgtcgg cgccggcggc gggatcgcct gtccgcgcgc cgcgctgctc 5160 gccttttccc tgggcgccgc ctttgtggtc accgggtccg tcaaccaact ggcccgcgag 5220 gctggcacca gcgacgcggt ccgactactg ctggcgacgg ccacctactc ggacgtggcc 5280 atggcgccgg gcggcgtcca ggtgctcaag aagcagacca tgttcgccgc gcgggccacg 5340 atgctcgccc agctgcaggc caagttcggc tcctttgacg ccgtgccgga gccgcagctg 5400 cgcaagctcg agcgctccgt gttcaagcag tccgtggcgg acgtgtgggc tgctgcacgc 5460 gaaaagtttg gtgtcgacgc taccgctgca agtccgcagg agaggatggc gctctgtgtg 5520 cgctggtaca tgtcgcagtc gtcgcgatgg gctaccgagg cgacgtccgc gcgcaaggcg 5580 gactaccaga tctggtgcgg ccccgccatc ggcagcttca acgacttcgt tcgcggcacc 5640 aagctggacg cgaccgctgg caccggcgag tttccgcgcg tcgtggacat caaccagcac 5700 atcctcctcg gagcctcgca ctaccgccgc gtgcagcaac aacaacagga cgacgacgta 5760 gaatacatca tcgtataa 5778 <21〇> 4 <211> 1925 <212> PRT <213> 裂殖壺菌物種 <4〇〇> 4 Met Pro Cys Asp Asn lie Ala Val Val Gly Met 15 10 Ala Val Gin Tyr Ala 1514 201038734 Ο ❹ aaggagcggc atcaacgact cgcgtgtcct tcggtctacc gtcgtggtgg cgctccgcca tactttgccg gacgacaagg gtgtcgccgc gtggagctcg gccgagtcgc gccatcggca ctcatcaaga cggccggtgg aagaacccgg tttggcgtgc gatgacgccg aaggtcaacg gacgacccag ggggagacgg ccggagaagc gccgggcgca gaccgcgtcg ctgcaccgcc gagaacggcg cagacggcct tgcctcaccg agcctcggcg cagctcacgc caggccttgc ggctacttgg ttcgtgcgcc gagtgtctgc ggcatgattg gagattttgg ctgcgcgggg gccgactttc gtggggccga tcgacccggc gcggcacctc cgcagtgggg gctgcctcga ccggcgtcga tcagccgaca gcgagggcag tctacgccag tactactcca ccgccgactc agctggagca gcgtgcgcgc cggcgctgtg cgccggtctc gcgagtcgcg tcctcacaga cccccaagct ccgagctggc ctgctgctgt tggctagtca tggagaagga actggatgtc cgttcatgta tgtggccggc actcgtggct ttgacgcgga actacgcgcg agatctccat agcgcctacg gcaagctatg ttcgcgccag tgctgatcgt gcgtgctgga ggcactgccc agattccgga gcagcaacag cgggcatcgt acaacatgcg cgcgttctcg gacgtcgtac ctttacgggc gctgggcaag cctctgtgcc ggaccaccct cggcgccctg tgtcgcgggc agtccacaac gggtcgccat agtgtccaag caacgtggga cctccacaac c gaggcgctg actggcggct cgagtacgcc catcgcgatc gctcctccga cgctttcact cggtgccacg gttggagctg gccatcgggc cggcgagggc tttgcacgag catgccgcgc ccagatcgag cgacgtgctc gctctttgcg cacctcgccg gaacgtgccg ccgcgccgaa caacgactcg gcgcctgggc cgaagtggcg cagccccgtc cagcatcacc cgacaaggtc ctccgccgcg cccgagcagg acgtcgtaca ccgtccttca ttcctgctcg accgccgaga cgcgccaact gtcctcaagc ctcacgtgcg gacgacaacg gcgccgcact ttgctcgcgc gacgtcgggt cgctacctcc tttacttgcc gtcgccagtg actcatgaga cgtggcgaca gcgcacgccg gctcatcgct gcgaccttgt gcagccaagg agcgcctttg cgcagcccct cgcatcaacg gcggtggatg atgttccgca ggggtgcagc ctgtcgcgac gtgtggtcga gcggacgccc atcgagaagg agcagcgcgc gggcggttgc ccctacacgc aagatgtaca gagttcgtgc agccgtgacg gtcagtgaca tgcaggagat tcggcaacct ccgtcaccga acacgcacca acctttacct ttgaggccag gccaggccga ccgcgcagcc agaagagggt tggccaactc accaggtgcc acgcctcggg cggccaaccc cgcgctcgcg cctccgagag gcagcaaccg ccgttgacga aaaccgggtc tgcgcttttt gtttggccct gtgtaccgcg cgccgacacc actacggcgt acaagaccgc ccgactcgca cgggcatctt ccaaggcgtg gcaactgcgg cacagctggc ccgtggagtc cgatcgggcc tgatcgccgg cgccgatgcc cgggcatcgc cctcggtcac agaagttgta gccacgatgt ttcttggcaa gatggactac cgtggccacg aggcgcaaac ggtggacgcc caaggcgcgc cgccgacggg cgttggctca cgctgaagcc ggtggagatg gccgctgagc gggctcggtg cgccgcgagc gcagtgggag tgcctggctg cgggtcctgc cctctcgctg tatcatggcc tgctactgac gcggctcgta gctgacaacg aagcgccaag tgtgaccagc cgggctcgac ggcgctgtgg gcgcgccgtg cgtgtccatc cttcggcctc cctgtcggac ggtggagttc cttctggcag cgacaaccgc caaacctgcc cgtcaagcaa gcacatccac caacgccgag cacgcgcatc cttcgtcgag ggctgccacc 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 15 201038734 ccgcatgtct ccgtggcgct ggaccgcccc agtgagtcgg cgtggacgca gaccctcaag 4140 tcgctggcgc tgctgaccgc ccaccgcgtg cccctgcaca acccgactct Gtttgcggac 4200 ctgtaccacc ccacgttcct gacggctatc gactctgcga tgcaggagcc cccgcccaag 4260 cccaaccgct tccttcgcag cgtagaggtc aacgggtact tttgccccg a cggcatcagc 4320 aagcaggttg ctgctgcaag tgccaaaccc tcgacgcatt gcatggttcg tttgcaccca 4380 gccaaggcag ttgtggttgc tgctgctggt gctgtggttg ctgattcgac gcccgtggtc 4440 aaggccaagc agacgtcgtc gtcgttgttg gttggggatg acgcctttct gcgctgctac 4500 gacgtggact ggccgctcta catgggcgcc atggcggaag gcatctcgtc ggtagacctg 4560 gtggtcgctg ccgccgaggc ccgcatgctg gcatcattcg gagcggcccg cttgcctatg 4620 gaccaggtgg aactccagat ccgtgagatc cagcaacgca cctccaacgc ctttgctgtc 4680 aacctgatgc cgggtcctga cgaggccgcg acggtggacg cgctgctgcg cacgggcgtc 47 40 tcaatcgtcg aggcatcggg ctacaccggc gcgctctctg cagacctggt gcgctaccgt 4800 gtcacgggtc tgcgacgaac tagttgcggt gcttctgtgt cggcgactca ccgtgtggtc 4860 gccaaggtgt cgcgcaccga ggtggccgag cactttctgc gcccggcgcc ggccgccgta 4920 ctagaggctt tggtcgccgc caaacagatt acgcccgagc aggccgcgct ggccagccgc 4980 gtcgccatgg ccgacgacgt cgcggtggag gccgactcgg gcgggcacac cgacaaccga 5040 ccgatccacg tgctgctgcc gctcgtggtg gcgcagcgca accgctggcg ccacctggtg 5100 gacacgccag tgcgcgtcgg cgccggcggc gggatcgcct gtccgcgcgc cgcgctgctc 5160 gccttttccc tgggcgccgc ctttgtggtc accgggtccg tcaaccaact ggcccgcgag 5220 gctggcacca gcgacgcggt ccgactactg ctggcgacgg ccacctactc ggacgtggcc 5280 atggcgccgg gcggcgtcca ggtgctcaag aagcagacca tgttcgccgc gcgggccacg 5340 atgctcgccc agctgcaggc caagttcggc tcctttgacg ccgtgccgga gccgcagctg 5400 cgcaagctcg agcgctccgt gttcaagcag tccgtggcgg acgtgtgggc tgctgcacgc 5460 gaaaagtttg gtgtcgacgc taccgctgca agtccgcagg agaggatggc gctctgtgtg 5520 cgctggtaca tgtcgcagtc gtcgcgatgg gctaccgagg cgacgtccgc gcgcaaggcg 5580 gactaccaga tctggtgcgg ccccgccatc ggcagcttca acgacttcgt tcgcggcacc 5640 aagctggacg cgaccgctgg caccggcgag tttccgcgcg tcgtggacat caaccagcac 5700 atcctcctcg gagcctcgca ctaccgccgc gtgcagcaac aacaacagga cgacgacgta 5760 gaatacatca tcgtataa 5778 < 21〇 > 4 < 211 > 1925 < 212 > PRT < 213 > Schizochytrium Bacterial species <4〇〇> 4 Met Pro Cys Asp Asn lie Ala Val Val Gly Met 15 10 Ala Val Gin Tyr Ala 15

Gly Cys Lys Asn Gin Asp Glu Phe Trp Asp Thr Leu Met Arg Lys Glu 20 25 30 16 201038734 工le Asn Ser Ser 35 Pro lie Ser Ala Glu Arg Leu Gly Thr Arq Tvr Ara 40 45 Asp Leu His Phe 50 His Pro Gin Arg Ser Lys Tyr Ala Asp Thr Phe Cvs 55 60 Asn Asp Arg Tyr 65 Gly Cys Val Asp Ala Ser Val Asp Asn Glu His Asp 70 75 80 Leu Leu Ala Asp Leu Ala Arg Arg Ala Leu Leu Asp Ala Gly lie Asn 85 90 95 Leu Asp Asp Ala 100 Ser Thr Thr Ala Asn Leu Arg Asp Phe Gly lie Val 105 110 Ser Gly Cys Leu 115 Ser Phe Pro Met Asp Asn Leu Gin Gly Glu Leu Leu 120 125 Asn Leu Tyr Gin 130 Val His Val Glu Asn Arg Val Gly Ala Gin Arg Phe 135 140 Arg Asp Ser Arg 145 Pro Trp Ser Glu Arg Pro Arg Ala Val Ser Pro Glu 150 155 160 Ala Ser Asp Pro Arg Val Tyr Ser Asp Pro Ala Ser Phe Val Ala Asn 165 170 175 Gin Leu Gly Leu • 180 Gly Pro Val Arg Tyr Ser Leu Asp Ala Ala Cys Ala 185 190 Ser Ala Leu Tyr 195 Cys Leu Lys Leu Ala Ser Asp His Leu Leu Ser Arg 200 205 Ser Ala Asp Val 210 Met Leu Cys Gly Ala Thr Cys Phe Pro Asp Pro Phe 215 220 Phe lie Leu Ser 〇 225 Gly Phe Ser Thr Phe Gin Ala Met Pro Leu Gly Gly 230 235 240 Pro Asp Asp Asn Pro Leu Ser Val Pro Leu Arg Gin Gly Ser Gin Gly 245 250 255Gly Cys Lys Asn Gin Asp Glu Phe Trp Asp Thr Leu Met Arg Lys Glu 20 25 30 16 201038734 Work Le Asn Ser Ser 35 Pro lie Ser Ala Glu Arg Leu Gly Thr Arq Tvr Ara 40 45 Asp Leu His Phe 50 His Pro Gin Arg Ser Lys Tyr Ala Asp Thr Phe Cvs 55 60 Asn Asp Arg Tyr 65 Gly Cys Val Asp Ala Ser Val Asp Asn Glu His Asp 70 75 80 Leu Leu Ala Asp Leu Ala Arg Arg Ala Leu Leu Asp Ala Gly lie Asn 85 90 95 Leu Asp Asp Ala 100 Ser Thr Thr Ala Asn Leu Arg Asp Phe Gly lie Val 105 110 Ser Gly Cys Leu 115 Ser Phe Pro Met Asp Asn Leu Gin Gly Glu Leu Leu 120 125 Asn Leu Tyr Gin 130 Val His Val Glu Asn Arg Val Gly Ala Gin Arg Phe 135 140 Arg Asp Ser Arg 145 Pro Trp Ser Glu Arg Pro Arg Ala Val Ser Pro Glu 150 155 160 Ala Ser Asp Pro Arg Val Tyr Ser Asp Pro Ala Ser Phe Val Ala Asn 165 170 175 Gin Leu Gly Leu • 180 Gly Pro Val Arg Tyr Ser Leu Asp Ala Ala Cys Ala 185 190 Ser Ala Leu Tyr 195 Cys Leu Lys Leu Ala Ser Asp His Leu Leu Ser Arg 200 205 Ser Ala Asp Val 210 Met Leu Cys Gly Ala Thr Cys Phe Pro Asp Pro Ph e 215 220 Phe lie Leu Ser 〇 225 Gly Phe Ser Thr Phe Gin Ala Met Pro Leu Gly Gly 230 235 240 Pro Asp Asp Asn Pro Leu Ser Val Pro Leu Arg Gin Gly Ser Gin Gly 245 250 255

Leu Thr Pro Gly Glu Gly Gly Ala lie Met Val Leu Lys Arg Leu Glu 260 265 270Leu Thr Pro Gly Glu Gly Gly Ala lie Met Val Leu Lys Arg Leu Glu 260 265 270

Asp Ala Val Arg 275 Asp Gly Asp Arg lie Tyr Gly Thr Leu Leu Gly Thr 280 285 Ser Leu Ser Asn 290 Ala Gly Cys Gly Leu Pro Leu Ser Pro His Leu Pro 295 300 Ser Glu Lys Ser 305 Cys Met Glu Asp Leu Tyr Thr Ser Val Gly lie Asp 310 315 320 Pro Ser Glu Val Gin Tyr Val Glu Cys His Ala Thr Gly Thr Pro Gin 325 330 335 17 201038734Asp Ala Val Arg 275 Asp Gly Asp Arg lie Tyr Gly Thr Leu Leu Gly Thr 280 285 Ser Leu Ser Asn 290 Ala Gly Cys Gly Leu Pro Leu Ser Pro His Leu Pro 295 300 Ser Glu Lys Ser 305 Cys Met Glu Asp Leu Tyr Thr Ser Val Gly lie Asp 310 315 320 Pro Ser Glu Val Gin Tyr Val Glu Cys His Ala Thr Gly Thr Pro Gin 325 330 335 17 201038734

Gly Asp Val Val Glu Val Glu Ala Leu Arg His Cys Phe Arg Gly Asn 340 345 350Gly Asp Val Val Glu Val Glu Ala Leu Arg His Cys Phe Arg Gly Asn 340 345 350

Thr Asp His Pro Pro Arg Met Gly Ser Thr Lys Gly Asn Phe Gly His 355 360 365Thr Asp His Pro Pro Arg Met Gly Ser Thr Lys Gly Asn Phe Gly His 355 360 365

Thr Leu Val Ala Ala Gly Phe Ala Gly Met Ala Lys Val Leu Leu Ser 370 375 380Thr Leu Val Ala Ala Gly Phe Ala Gly Met Ala Lys Val Leu Leu Ser 370 375 380

Met Gin His Gly Thr lie Pro Pro Thr Pro Gly Val Asp Arg Ser Asn 385 390 395 400Met Gin His Gly Thr lie Pro Pro Thr Pro Gly Val Asp Arg Ser Asn 385 390 395 400

Cys lie Asp Pro Leu Val Val Asp Glu Ala lie Pro Trp Pro Tyr Ser 405 410 415Cys lie Asp Pro Leu Val Val Asp Glu Ala lie Pro Trp Pro Tyr Ser 405 410 415

Ser Ala Gin Ala Arg Ala Gly Lys Pro Gly Asp Glu Leu Lys Cys Ala 420 425 430Ser Ala Gin Ala Arg Ala Gly Lys Pro Gly Asp Glu Leu Lys Cys Ala 420 425 430

Ser Leu Ser Ala Phe Gly Phe Gly Gly Thr Asn Ala His Cys Val Phe 435 440 445Ser Leu Ser Ala Phe Gly Phe Gly Gly Thr Asn Ala His Cys Val Phe 435 440 445

Arg Glu His Arg Gin 工le Ala Ala Thr Ala Thr Ala Ser Pro Val Leu 450 455 460Arg Glu His Arg Gin work le Ala Ala Thr Ala Thr Ala Ser Pro Val Leu 450 455 460

Pro Glu Val Thr Pro Gly Pro 工le Ala lie lie Gly Met Asp Ala Thr 465 470 475 480Pro Glu Val Thr Pro Gly Pro Ale lie lie Gly Met Asp Ala Thr 465 470 475 480

Phe Gly Thr Leu Lys Gly Leu Asp Ala Phe Glu Gin Ala lie Tyr Lys 485 490 495Phe Gly Thr Leu Lys Gly Leu Asp Ala Phe Glu Gin Ala lie Tyr Lys 485 490 495

Gly Thr Asp Gly Ala Ser Asp Leu Pro Ser Lys Arg Trp Arg Phe Leu 500 505 510Gly Thr Asp Gly Ala Ser Asp Leu Pro Ser Lys Arg Trp Arg Phe Leu 500 505 510

Gly Ala Asp Thr Asp Phe Leu Thr Ala Met Gly Leu Asp Ala Val Pro 515 520 525Gly Ala Asp Thr Asp Phe Leu Thr Ala Met Gly Leu Asp Ala Val Pro 515 520 525

Arg Gly Cys Tyr Val Arg Asp Val Asp Val Asp Tyr Lys Arg Leu Arg 530 535 540Arg Gly Cys Tyr Val Arg Asp Val Asp Val Asp Tyr Lys Arg Leu Arg 530 535 540

Ser Pro Met lie Pro Glu Asp Val Leu Arg Pro Gin Gin Leu Leu Ala 545 550 555 560Ser Pro Met lie Pro Glu Asp Val Leu Arg Pro Gin Gin Leu Leu Ala 545 550 555 560

Val Ala Thr Met Asp Arg Ala Leu Gin Asp Ala Gly Met Ala Thr Gly 565 570 575Val Ala Thr Met Asp Arg Ala Leu Gin Asp Ala Gly Met Ala Thr Gly 565 570 575

Gly Lys Val Ala Val Leu Val Gly Leu Gly Thr Asp Thr Glu Leu Tyr 580 585 590Gly Lys Val Ala Val Leu Val Gly Leu Gly Thr Asp Thr Glu Leu Tyr 580 585 590

Arg His Arg Ala Arg Val Thr Leu Lys Glu Arg Leu Asp Pro Ala Ala 595 600 605Arg His Arg Ala Arg Val Thr Leu Lys Glu Arg Leu Asp Pro Ala Ala 595 600 605

Phe Ser Pro Glu Gin Val Gin Glu Met Met Asp Tyr lie Asn Asp Cys 610 615 620Phe Ser Pro Glu Gin Val Gin Glu Met Met Asp Tyr lie Asn Asp Cys 610 615 620

Gly Thr Ser Thr Ser Tyr Thr Ser Tyr lie Gly Asn Leu Val Ala Thr 18 201038734 625 630 635 640Gly Thr Ser Thr Ser Tyr Thr Ser Tyr lie Gly Asn Leu Val Ala Thr 18 201038734 625 630 635 640

Arg Val Ser Ser Gin Trp Gly Phe Thr Gly Pro Ser Phe Thr Val Thr 645 650 655Arg Val Ser Ser Gin Trp Gly Phe Thr Gly Pro Ser Phe Thr Val Thr 645 650 655

Glu Gly Ala Asn Ser Val Tyr Arg Cys Leu Glu Leu Gly Lys Phe Leu 660 665 67 0Glu Gly Ala Asn Ser Val Tyr Arg Cys Leu Glu Leu Gly Lys Phe Leu 660 665 67 0

Leu Asp Thr His Gin Val Asp Ala Val Val Val Ala Gly Val Asp Leu 675 680 685Leu Asp Thr His Gin Val Asp Ala Val Val Val Ala Gly Val Asp Leu 675 680 685

Cys Ala Thr Ala Glu Asn Leu Tyr Leu Lys Ala Arg Arg Ser Ala lie 690 695 700Cys Ala Thr Ala Glu Asn Leu Tyr Leu Lys Ala Arg Arg Ser Ala lie 690 695 700

Ser Arg Gin Asp His Pro Arg Ala Asn Phe Glu Ala Ser Ala Asp Gly 705 710 715 720 ❹Ser Arg Gin Asp His Pro Arg Ala Asn Phe Glu Ala Ser Ala Asp Gly 705 710 715 720 ❹

Tyr Phe Ala Gly Glu Gly Ser Gly Ala Leu Val Leu Lys Arg Gin Ala 725 730 735Tyr Phe Ala Gly Glu Gly Ser Gly Ala Leu Val Leu Lys Arg Gin Ala 725 730 735

Asp Val Gly Ser Asp Asp Lys Val Tyr Ala Ser Val Ala Gly Leu Thr 740 745 750Asp Val Gly Ser Asp Asp Lys Val Tyr Ala Ser Val Ala Gly Leu Thr 740 745 750

Cys Ala Ala Gin Pro Ala Glu Ala Val Ser Pro Leu Leu Leu Gin Val 755 760 765Cys Ala Ala Gin Pro Ala Glu Ala Val Ser Pro Leu Leu Leu Gin Val 755 760 765

His Asn Asp Asp Asn Glu Lys Arg Val Val Glu Met Val Glu Leu Ala 770 775 780His Asn Asp Asp Asn Glu Lys Arg Val Val Glu Met Val Glu Leu Ala 770 775 780

Ala Asp Ser Gly Arg His Ala Pro His Leu Ala Asn Ser Pro Leu Ser 785 79〇 795 800Ala Asp Ser Gly Arg His Ala Pro His Leu Ala Asn Ser Pro Leu Ser 785 79〇 795 800

Ala Glu Ser Gin Leu Glu Gin Val Ser Lys Leu Leu Ala His Gin Val 805 810 815Ala Glu Ser Gin Leu Glu Gin Val Ser Lys Leu Leu Ala His Gin Val 805 810 815

Pro Gly Ser Val Ala lie Gly Ser Val Arg Ala Asn Val Gly Asp Val 820 825 830Pro Gly Ser Val Ala lie Gly Ser Val Arg Ala Asn Val Gly Asp Val 820 825 830

Gly Tyr Ala Ser Gly Ala Ala Ser Leu lie Lys Thr Ala Leu Cys Leu 835 840 845Gly Tyr Ala Ser Gly Ala Ala Ser Leu lie Lys Thr Ala Leu Cys Leu 835 840 845

His Asn Arg Tyr Leu Pro Ala Asn Pro Gin Trp Glu Arg Pro Val Ala 850 855 860His Asn Arg Tyr Leu Pro Ala Asn Pro Gin Trp Glu Arg Pro Val Ala 850 855 860

Pro Val Ser Glu Ala Leu Phe Thr Cys Pro Arg Ser Arg Ala Trp Leu 865 870 875 880Pro Val Ser Glu Ala Leu Phe Thr Cys Pro Arg Ser Arg Ala Trp Leu 865 870 875 880

Lys Asn Pro Gly Glu Ser Arg Leu Ala Ala Val Ala Ser Ala Ser Glu 885 890 895Lys Asn Pro Gly Glu Ser Arg Leu Ala Ala Val Ala Ser Ala Ser Glu 885 890 895

Ser Gly Ser Cys Phe Gly Val Leu Leu Thr Asp Glu Tyr Ala Thr His 900 905 910Ser Gly Ser Cys Phe Gly Val Leu Leu Thr Asp Glu Tyr Ala Thr His 900 905 910

Glu Ser Ser Asn Arg Leu Ser Leu Asp Asp Ala Ala Pro Lys Leu lie 915 920 925 19 201038734Glu Ser Ser Asn Arg Leu Ser Leu Asp Asp Ala Ala Pro Lys Leu lie 915 920 925 19 201038734

Ala lie Arg Gly Asp Thr Val Asp Asp lie Met Ala Lys Val Asn Ala 930 935 940Ala lie Arg Gly Asp Thr Val Asp Asp lie Met Ala Lys Val Asn Ala 930 935 940

Glu Leu Ala Leu Leu Arg Ala His Ala Glu Thr Gly Ser Ala Thr Asp 945 950 955 960Glu Leu Ala Leu Leu Arg Ala His Ala Glu Thr Gly Ser Ala Thr Asp 945 950 955 960

Asp Asp Pro Ala Ala Ala Val Ala Phe Thr Ala His Arg Leu Arg Phe 965 970 975Asp Asp Pro Ala Ala Ala Val Ala Phe Thr Ala His Arg Leu Arg Phe 965 970 975

Leu Arg Leu Val Gly Glu Thr Val Ala Ser His Gly Ala Thr Ala Thr 980 985 990Leu Arg Leu Val Gly Glu Thr Val Ala Ser His Gly Ala Thr Ala Thr 980 985 990

Leu Cys Leu Ala Leu Leu Thr Thr Pro Glu Lys Leu Glu Lys Glu Leu 995 1000 1005Leu Cys Leu Ala Leu Leu Thr Thr Pro Glu Lys Leu Glu Lys Glu Leu 995 1000 1005

Glu Leu Ala Ala Lys Gly Val Pro Arg Ser Ala Lys Ala Gly Arg 1010 1015 1020Glu Leu Ala Ala Lys Gly Val Pro Arg Ser Ala Lys Ala Gly Arg 1010 1015 1020

Asn Trp Met Ser Pro Ser Gly Ser Ala Phe Ala Pro Thr Pro Val 1025 1030 1035Asn Trp Met Ser Pro Ser Gly Ser Ala Phe Ala Pro Thr Pro Val 1025 1030 1035

Thr Ser Asp Arg Val Ala Phe Met Tyr Gly Glu Gly Arg Ser Pro 1040 1045 1050Thr Ser Asp Arg Val Ala Phe Met Tyr Gly Glu Gly Arg Ser Pro 1040 1045 1050

Tyr Tyr Gly Val Gly Leu Asp Leu His Arg Leu Trp Pro Ala Leu 1055 1060 1065Tyr Tyr Gly Val Gly Leu Asp Leu His Arg Leu Trp Pro Ala Leu 1055 1060 1065

His Glu Arg lie Asn Asp Lys Thr Ala Ala Leu Trp Glu Asn Gly 1070 1075 1080His Glu Arg lie Asn Asp Lys Thr Ala Ala Leu Trp Glu Asn Gly 1070 1075 1080

Asp Ser Trp Leu Met Pro Arg Ala Val Asp Ala Asp Ser Gin Arg 1085 1090 1095Asp Ser Trp Leu Met Pro Arg Ala Val Asp Ala Asp Ser Gin Arg 1085 1090 1095

Ala Val Gin Thr Ala Phe Asp Ala Asp Gin lie Glu Met Phe Arg 1100 1105 1110 iiAla Val Gin Thr Ala Phe Asp Ala Asp Gin lie Glu Met Phe Arg 1100 1105 1110 ii

Thr Gly lie Phe Val Ser lie Cys Leu Thr Asp Tyr Ala Arg Asp 1115 1120 1125Thr Gly lie Phe Val Ser lie Cys Leu Thr Asp Tyr Ala Arg Asp 1115 1120 1125

Val Leu Gly Val Gin Pro Lys Ala Cys Phe Gly Leu Ser Leu Gly 1130 1135 1140Val Leu Gly Val Gin Pro Lys Ala Cys Phe Gly Leu Ser Leu Gly 1130 1135 1140

Glu lie Ser Met Leu Phe Ala Leu Ser Arg Arg Asn Cys Gly Leu 1145 1150 1155Glu lie Ser Met Leu Phe Ala Leu Ser Arg Arg Asn Cys Gly Leu 1145 1150 1155

Ser Asp Gin Leu Thr Gin Arg Leu Arg Thr Ser Pro Val Trp Ser 1160 1165 1170Ser Asp Gin Leu Thr Gin Arg Leu Arg Thr Ser Pro Val Trp Ser 1160 1165 1170

Thr Gin Leu Ala Val Glu Phe Gin Ala Leu Arg Lys Leu Trp Asn 1175 1180 1185Thr Gin Leu Ala Val Glu Phe Gin Ala Leu Arg Lys Leu Trp Asn 1175 1180 1185

Val Pro Ala Asp Ala Pro Val Glu Ser Phe Trp Gin Gly Tyr Leu 1190 1195 1200Val Pro Ala Asp Ala Pro Val Glu Ser Phe Trp Gin Gly Tyr Leu 1190 1195 1200

Val Arg Ala Ser Arg Ala Glu lie Glu Lys Ala lie Gly Pro Asp 1205 1210 1215 20 201038734Val Arg Ala Ser Arg Ala Glu lie Glu Lys Ala lie Gly Pro Asp 1205 1210 1215 20 201038734

Asn Arg Phe Val Arg Leu Leu lie Val Asn Asp Ser Ser Ser Ala 1220 1225 1230Asn Arg Phe Val Arg Leu Leu lie Val Asn Asp Ser Ser Ser Ala 1220 1225 1230

Leu lie Ala Gly Lys Pro Ala Glu Cys Leu Arg Val Leu Glu Arg 1235 1240 1245 Leu Gly Gly Arg Leu Pro Pro Met Pro Val Lys Gin Gly Met lie 1250 1255 1260 Gly His Cys Pro Glu Val Ala Pro Tyr Thr Pro Gly lie Ala His 1265 1270 1275 lie His Glu lie Leu Glu lie Pro Asp Ser Pro Val Lys Met Tyr 1280 1285 1290 Thr Ser Val Thr Asn Ala Glu Leu Arg Gly Gly Ser Asn Ser Ser 1295 1300 1305 O lie Thr Glu Phe Val Gin Lys Leu Tyr Thr Arg lie Ala Asp Phe 1310 1315 1320 Pro Gly lie Val Asp Lys Val Ser Arg Asp Gly His Asp Val Phe 1325 1330 1335 Val Glu Val Gly Pro Asn Asn Met Arg Ser Ala Ala Val Ser Asp 1340 1345 1350 . He Leu Gly Lys Ala Ala Thr Pro His Val Ser Val Ala Leu Asp 1355 1360 1365 Arg Pro Ser Glu Ser Ala Trp Thr Gin Thr Leu Lys Ser Leu Ala 1370 1375 1380 Leu Leu Thr Ala His Arg Val Pro Leu His Asn Pro Thr Leu Phe 1385 1390 1395 一 Ala 〇 Asp Leu Tyr His Pro Thr Phe Leu Thr Ala lie Asp Ser Ala 1400 1405 1410 Met Gin Glu Pro Pro Pro Lys Pro Asn Arg Phe Leu Arg Ser Val 1415 1420 1425 Glu Val Asn Gly Tyr Phe Cys Pro Asp Gly lie Ser Lys Gin Val 1430 1435 1440 Ala Ala Ala Ser Ala Lys Pro Ser Thr His Cys Met Val Arg Leu 1445 1450 1455 His Pro Ala Lys Ala Val Val Val Ala Ala Ala Gly Ala Val Val 1460 1465 1470 Ala Asp Ser Thr Pro Val Val Lys Ala Lys Gin Thr Ser Ser Ser 1475 1480 1485Leu lie Ala Gly Lys Pro Ala Glu Cys Leu Arg Val Leu Glu Arg 1235 1240 1245 Leu Gly Gly Arg Leu Pro Pro Met Pro Val Lys Gin Gly Met lie 1250 1255 1260 Gly His Cys Pro Glu Val Ala Pro Tyr Thr Pro Gly lie Ala His 1265 1270 1275 lie His Glu lie Leu Glu lie Pro Asp Ser Pro Val Lys Met Tyr 1280 1285 1290 Thr Ser Val Thr Asn Ala Glu Leu Arg Gly Gly Ser Asn Ser Ser 1295 1300 1305 O lie Thr Glu Phe Val Gin Lys Leu Tyr Thr Arg lie Ala Asp Phe 1310 1315 1320 Pro Gly lie Val Asp Lys Val Ser Arg Asp Gly His Asp Val Phe 1325 1330 1335 Val Glu Val Gly Pro Asn Asn Met Arg Ser Ala Ala Val Ser Asp 1340 1345 1350 . He Leu Gly Lys Ala Ala Thr Pro His Val Ser Val Ala Leu Asp 1355 1360 1365 Arg Pro Ser Glu Ser Ala Trp Thr Gin Thr Leu Lys Ser Leu Ala 1370 1375 1380 Leu Leu Thr Ala His Arg Val Pro Leu His Asn Pro Thr Leu Phe 1385 1390 1395 Ala 〇Asp Leu Tyr His Pro Thr Phe Leu Thr Ala lie Asp Ser Ala 1400 1405 1410 Met Gin Glu Pro Pro Pro Lys Pro Asn Arg Phe Leu Arg Ser Val 1415 1420 1425 G Lu Val Asn Gly Tyr Phe Cys Pro Asp Gly lie Ser Lys Gin Val 1430 1435 1440 Ala Ala Ala Ser Ala Lys Pro Ser Thr His Cys Met Val Arg Leu 1445 1450 1455 His Pro Ala Lys Ala Val Val Val Ala Ala Ala Gly Ala Val Val 1460 1465 1470 Ala Asp Ser Thr Pro Val Val Lys Ala Lys Gin Thr Ser Ser Ser 1475 1480 1485

Leu Leu Val Gly Asp Asp Ala Phe Leu Arg Cys Tyr Asp Val Asp 1490 1495 1500 21 201038734Leu Leu Val Gly Asp Asp Ala Phe Leu Arg Cys Tyr Asp Val Asp 1490 1495 1500 21 201038734

Trp Pro Leu Tyr Met Gly Ala Met Ala Glu Gly lie Ser Ser Val 1505 1510 1515Trp Pro Leu Tyr Met Gly Ala Met Ala Glu Gly lie Ser Ser Val 1505 1510 1515

Asp Leu Val Val Ala Ala Ala Glu Ala Arg Met Leu Ala Ser Phe 1520 1525 1530Asp Leu Val Val Ala Ala Ala Glu Ala Arg Met Leu Ala Ser Phe 1520 1525 1530

Gly Ala Ala Arg Leu Pro Met Asp Gin Val Glu Leu Gin lie Arg 1535 1540 1545Gly Ala Ala Arg Leu Pro Met Asp Gin Val Glu Leu Gin lie Arg 1535 1540 1545

Glu lie Gin Gin Arg Thr Ser Asn Ala Phe Ala Val Asn Leu Met 1550 1555 1560Glu lie Gin Gin Arg Thr Ser Asn Ala Phe Ala Val Asn Leu Met 1550 1555 1560

Pro Gly Pro Asp Glu Ala Ala Thr Val Asp Ala Leu Leu Arg Thr 1565 1570 1575Pro Gly Pro Asp Glu Ala Ala Thr Val Asp Ala Leu Leu Arg Thr 1565 1570 1575

Gly Val Ser lie Val Glu Ala Ser Gly Tyr Thr Gly Ala Leu Ser 1580 1585 1590Gly Val Ser lie Val Glu Ala Ser Gly Tyr Thr Gly Ala Leu Ser 1580 1585 1590

Ala Asp Leu Val Arg Tyr Arg Val Thr Gly Leu Arg Arg Thr Ser 1595 1600 1605Ala Asp Leu Val Arg Tyr Arg Val Thr Gly Leu Arg Arg Thr Ser 1595 1600 1605

Cys Gly Ala Ser Val Ser Ala Thr His Arg Val Val Ala Lys Val 1610 1615 1620Cys Gly Ala Ser Val Ser Ala Thr His Arg Val Val Ala Lys Val 1610 1615 1620

Ser Arg Thr Glu Val Ala Glu His Phe Leu Arg Pro Ala Pro Ala 1625 1630 1635Ser Arg Thr Glu Val Ala Glu His Phe Leu Arg Pro Ala Pro Ala 1625 1630 1635

Ala Val Leu Glu Ala Leu Val Ala Ala Lys Gin lie Thr Pro Glu 1640 1645 1650Ala Val Leu Glu Ala Leu Val Ala Ala Lys Gin lie Thr Pro Glu 1640 1645 1650

Gin Ala Ala Leu Ala Ser Arg Val Ala Met Ala Asp Asp Val Ala 1655 1660 1665Gin Ala Ala Leu Ala Ser Arg Val Ala Met Ala Asp Asp Val Ala 1655 1660 1665

Val Glu Ala Asp Ser Gly Gly His Thr Asp Asn Arg Pro lie His 1670 1675 1680Val Glu Ala Asp Ser Gly Gly His Thr Asp Asn Arg Pro lie His 1670 1675 1680

Val Leu Leu Pro Leu Val Val Ala Gin Arg Asn Arg Trp Arg His 1685 1690 1695Val Leu Leu Pro Leu Val Val Ala Gin Arg Asn Arg Trp Arg His 1685 1690 1695

Leu Val Asp Thr Pro Val Arg Val Gly Ala Gly Gly Gly lie Ala 1700 1705 1710Leu Val Asp Thr Pro Val Arg Val Gly Ala Gly Gly Gly lie Ala 1700 1705 1710

Cys Pro Arg Ala Ala Leu Leu Ala Phe Ser Leu Gly Ala Ala Phe 1715 1720 1725Cys Pro Arg Ala Ala Leu Leu Ala Phe Ser Leu Gly Ala Ala Phe 1715 1720 1725

Val Val Thr Gly Ser Val Asn Gin Leu Ala Arg Glu Ala Gly Thr 1730 1735 1740Val Val Thr Gly Ser Val Asn Gin Leu Ala Arg Glu Ala Gly Thr 1730 1735 1740

Ser Asp Ala Val Arg Leu Leu Leu Ala Thr Ala Thr Tyr Ser Asp 1745 1750 1755Ser Asp Ala Val Arg Leu Leu Leu Ala Thr Ala Thr Tyr Ser Asp 1745 1750 1755

Val Ala Met Ala Pro Gly Gly Val Gin Val Leu Lys Lys Gin Thr 1760 1765 1770Val Ala Met Ala Pro Gly Gly Val Gin Val Leu Lys Lys Gin Thr 1760 1765 1770

Met Phe Ala Ala Arg Ala Thr Met Leu Ala Gin Leu Gin Ala Lys 22 201038734 1775 1780 1785Met Phe Ala Ala Arg Ala Thr Met Leu Ala Gin Leu Gin Ala Lys 22 201038734 1775 1780 1785

Phe Gly Ser Phe Asp Ala Val Pro Glu Pro Gin Leu Arg Lys Leu 1790 1795 1800Phe Gly Ser Phe Asp Ala Val Pro Glu Pro Gin Leu Arg Lys Leu 1790 1795 1800

Glu Arg Ser Val Phe Lys Gin Ser Val Ala Asp Val Trp Ala Ala 1805 1810 1815Glu Arg Ser Val Phe Lys Gin Ser Val Ala Asp Val Trp Ala Ala 1805 1810 1815

Ala Arg Glu Lys Phe Gly Val Asp Ala Thr Ala Ala Ser Pro Gin 1820 1825 1830Ala Arg Glu Lys Phe Gly Val Asp Ala Thr Ala Ala Ser Pro Gin 1820 1825 1830

Glu Arg Met Ala Leu Cys Val Arg Trp Tyr Met Ser Gin Ser Ser 1835 1840 1845Glu Arg Met Ala Leu Cys Val Arg Trp Tyr Met Ser Gin Ser Ser 1835 1840 1845

Arg Trp Ala Thr Glu Ala Thr Ser Ala Arg Lys Ala Asp Tyr Gin 1850 1855 I860Arg Trp Ala Thr Glu Ala Thr Ser Ala Arg Lys Ala Asp Tyr Gin 1850 1855 I860

lie Trp Cys Gly Pro Ala lie Gly Ser Phe Asn Asp Phe Val Arg 1865 1870 1875Lie Trp Cys Gly Pro Ala lie Gly Ser Phe Asn Asp Phe Val Arg 1865 1870 1875

Gly Thr Lys Leu Asp Ala Thr Ala Gly Thr Gly Glu Phe Pro Arg 1880 1885 1890Gly Thr Lys Leu Asp Ala Thr Ala Gly Thr Gly Glu Phe Pro Arg 1880 1885 1890

Val Val Asp lie _Asn Gin His lie Leu Leu Gly Ala Ser His Tyr 1895 1900 1905Val Val Asp lie _Asn Gin His lie Leu Leu Gly Ala Ser His Tyr 1895 1900 1905

Arg Arg Val Gin Gin Gin Gin Gin Asp Asp Asp Val Glu Tyr lie 1910 1915 1920Arg Arg Val Gin Gin Gin Gin Gin Asp Asp Asp Val Glu Tyr lie 1910 1915 1920

He Val 1925 <210> 5 <211> 4308 <212> DNA <213> 裂殖壺菌物種 <400> 5 60 120 180 240 300 360 420 480 540 600 660 720 atgacatcat cgaagaagac tcccgtgtgg gagatgagca aggaggagct gctggacggc aagacggtgg tcttcgacta caacgagctg ctcgaattcg ccgagggcga cgtgggccaa gtgttcggac ccgagttcga catcatcgac aagtaccggc gtcgcgtgcg gctgccggcg cgcgagtacc tgctcgtgtc gcgcgtgacg ctgatggacg ccgaggtgaa caacttccgc gtcgggtcgc gcatggtgac cgagtacgac gtgcccgtga acggggagct gtcggagggc ggggacgtgc cgtgggcggt gctggtggag tcggggcagt gcgacctgat gctcatctcg tacatgggca tcgacttcca gtgcaagggc gaccgcgtgt accgcctgct caacacatcg ctcaccttct tcggggtggc gcacgagggc gagacgctgg tgtacgacat ccgcgtcacg gggttcgcca agggcgcggg cggggagatc tcgatgttct tcttcgagta cgactgcttc gtggacggcc gcctgctgat cgagatgcgc gacgggtgcg ccgggttctt cacggacgcc gagctggccg ccggcaaggg cgtgcttaag accaaggcgg agctggcggc gcgcgcgcag atccagaagc aggacatcgc gccctttgcg ccggcgccgt gctcgcacaa gacctcgctg gacgcgcgcg agatgcggct gctcgtggac cgccagtggg cgcgcgtctt cggcagcggc 780 23 201038734 atggcgggca tcgactacaa gttgtgcgct cgcaagatgc tcatgatcga ccgcgtcacg 840 cacctcgacc cgcgcggcgg cgcgcacggc ctcgggctgc tgatcgggga gaaggtgctg 900 gagcgcgacc actggtactt cccctgccac tttgtgcgcg acgaggtgat ggccgggtcg 960 ctggtcagcg acggctgctc gcagctcctc aaggtgtaca tgctgtggct cggcctgcac 1020 acgaccgtgg gcgcgttcga ctttcgtccc gtgagcgggc acgccaacaa ggtgcggtgc 1080 cgcgggcaga tctcaccgca caagggcaag ctcgtgtacg tgatggagat caaggaaatg 1140 ggctttgacg cgaagacggg cgatccgttt gcgatcgcgg acgtggacat catcgacgtc 1200 aacttcgagg agggacaggc gtttgcggga gtggaagacc tgcacagcta cggccagggc 1260 gacctccgca agaagatcgt cgtcgacttc aagggcatcg cgctctccct gcagaagcgg 1320 aaggagcagc agaaggaaag catgaccgtg actacgacga cgacgacgac gagccgggtg 1380 attgcgccgc ccagcgggtg cctcaagggc gacccgacgg cgccgacgag cgtgacgtgg 1440 cacccgatgg cggagggcaa cggcgggccc ggaccgacgc cgtcgttctc gccgtccgcg 1500 tacccgccgc gggcggtgtg cttctcgccg ttccccaaca acccgcttga caacgaccac 1560 acgccgggcc agatgccgtt gacctggttc aacatgtccg aattcatgtg cggcaaagtg 1620 tccaactgcc tgggccccga gtttgcgcgc ttcgacgcga gcaagacgag ccgcagcccg 1680 gcctttgacc tggcgctcgt gacgcgggtg acgagcgtgg cggacatgga gcacgggccg 17 40 ttctacaacg tggacgtcaa cccgggccag ggcacgatgg tgggcgagtt cgactgtccc 1800 gcggacgcgt ggttcttcgg cgcctcgagc cgcgacgacc acatgccgta ctcgatcctg I860 atggagatcg cgctgcagac gtcgggcgtc ctcacctcgg tgctcaaggc gccgctgacg 1920 atggacaagg acgacatcct cttccgcaac ctcgacgcag acgccgagct cgtgggcgac 1980 gccatgccgg acgtgcgcgg caagacgatc cgcaacttca ccaagtgcac aggctacagc 2040 atgctcggca agatgggcat ccaccgcttc acctttgagc tcagcgtcga cggcgccgtc 2100 ttctacaagg gcagcacctc gtttggctgg ttcgtccccg aggtcttcga gtcgcagacc 2160 ggtctcgaca acggcaagcc gcgcctgcct tggtaccgcg agaacaacgt cgccgtcgac 2220 acgctctccg cgcccgcctc cgcttcctcc gcgcaaggtc agctgcagct gcagcgacgc 2280 gggtcgcagg cgcagttcct ggacacaatc cacctggcgg gcagcggcgc cggcgtgcac 2340 ggccagggct acgcgcacgg ggagaaggcc gtgaacaagc aagattggtt cttctcgtgc 2400 cacttctggt tcgaccccgt gatgcccggg tccctgggca tcgagtcgat gttccagctc 2460 gtcgaggcgt ggtgcgtgaa gcagggactc gcggcgcggc acggcatcgc tcacccagtg 2520 ttcgcgcacg cgcccggggc cacgagctgg aagtaccgcg ggcagctaac ccccaagaac 2580 gaccgcatgg acagcgaggt gcacatcaag tcggtggcgg ccttctcctc ctgggtcgac 2640 gtcgtcgcgg acgggttcct cttcgtcgac ggcctccgcg tctactcggc agacaacctc 2700 cgcgtccgca tccagaccgg cgccggccac gttgaagagc aagaggttgc tgccaaggcc 2760 acaaccaaga acagcagtat tgctgatgtg gacgtggcgg acctgcaagc gctcaagcag 2820 gcgttgctga cgctggagcg accgctgcag ctggacgcgg ggagcgaggt gcccgcctgc 2880 gcggtgagcg acctgggcga taggggcttc atggagacgt acggggtggt ggcgccgctg 2940 tacagcgggg cgatggccaa gggcatcgcg tcggcggacc tggtgatcgc gatgggccag 3000He Val 1925 <210> 5 <211> 4308 <212> DNA <213> Schizochytrium species <400> 5 60 120 180 240 300 360 420 480 540 600 660 720 atgacatcat cgaagaagac tcccgtgtgg gagatgagca aggaggagct gctggacggc aagacggtgg tcttcgacta caacgagctg ctcgaattcg ccgagggcga cgtgggccaa gtgttcggac ccgagttcga catcatcgac aagtaccggc gtcgcgtgcg gctgccggcg cgcgagtacc tgctcgtgtc gcgcgtgacg ctgatggacg ccgaggtgaa caacttccgc gtcgggtcgc gcatggtgac cgagtacgac gtgcccgtga acggggagct gtcggagggc ggggacgtgc cgtgggcggt gctggtggag tcggggcagt gcgacctgat gctcatctcg tacatgggca tcgacttcca gtgcaagggc gaccgcgtgt accgcctgct caacacatcg ctcaccttct tcggggtggc gcacgagggc gagacgctgg tgtacgacat ccgcgtcacg gggttcgcca agggcgcggg cggggagatc tcgatgttct tcttcgagta cgactgcttc gtggacggcc gcctgctgat Cgagatgcgc gacgggtgcg ccgggttctt cacggacgcc gagctggccg ccggcaaggg cgtgcttaag accaaggcgg agctggcggc gcgcgcgca atccagaagc aggacatcgc gccctttgcg ccggcgccgt gctcgcacaa gacctcgctg gacgcgcgcg agatgcggct gctcgtggac cgccagtggg cgcgcctctt cg gcagcggc 780 23 201038734 atggcgggca tcgactacaa gttgtgcgct cgcaagatgc tcatgatcga ccgcgtcacg 840 cacctcgacc cgcgcggcgg cgcgcacggc ctcgggctgc tgatcgggga gaaggtgctg 900 gagcgcgacc actggtactt cccctgccac tttgtgcgcg acgaggtgat ggccgggtcg 960 ctggtcagcg acggctgctc gcagctcctc aaggtgtaca tgctgtggct cggcctgcac 1020 acgaccgtgg gcgcgttcga ctttcgtccc gtgagcgggc acgccaacaa ggtgcggtgc 1080 cgcgggcaga tctcaccgca caagggcaag ctcgtgtacg tgatggagat caaggaaatg 1140 ggctttgacg cgaagacggg cgatccgttt gcgatcgcgg acgtggacat catcgacgtc 1200 aacttcgagg agggacaggc gtttgcggga gtggaagacc tgcacagcta cggccagggc 1260 gacctccgca agaagatcgt cgtcgacttc aagggcatcg cgctctccct gcagaagcgg 1320 aaggagcagc agaaggaaag catgaccgtg actacgacga cgacgacgac gagccgggtg 1380 attgcgccgc ccagcgggtg cctcaagggc gacccgacgg cgccgacgag cgtgacgtgg 1440 cacccgatgg cggagggcaa cggcgggccc ggaccgacgc cgtcgttctc gccgtccgcg 1500 tacccgccgc gggcggtgtg cttctcgccg ttccccaaca acccgcttga caacgaccac 1560 acgccgggcc agatgccgtt gacctggttc aacatgtc cg aattcatgtg cggcaaagtg 1620 tccaactgcc tgggccccga gtttgcgcgc ttcgacgcga gcaagacgag ccgcagcccg 1680 gcctttgacc tggcgctcgt gacgcgggtg acgagcgtgg cggacatgga gcacgggccg 17 40 ttctacaacg tggacgtcaa cccgggccag ggcacgatgg tgggcgagtt cgactgtccc 1800 gcggacgcgt ggttcttcgg cgcctcgagc cgcgacgacc acatgccgta ctcgatcctg I860 atggagatcg cgctgcagac gtcgggcgtc ctcacctcgg tgctcaaggc gccgctgacg 1920 atggacaagg acgacatcct cttccgcaac ctcgacgcag acgccgagct cgtgggcgac 1980 gccatgccgg acgtgcgcgg caagacgatc cgcaacttca ccaagtgcac aggctacagc 2040 atgctcggca agatgggcat ccaccgcttc acctttgagc tcagcgtcga cggcgccgtc 2100 ttctacaagg gcagcacctc gtttggctgg ttcgtccccg aggtcttcga gtcgcagacc 2160 ggtctcgaca acggcaagcc gcgcctgcct tggtaccgcg agaacaacgt cgccgtcgac 2220 acgctctccg cgcccgcctc cgcttcctcc gcgcaaggtc agctgcagct gcagcgacgc 2280 gggtcgcagg cgcagttcct ggacacaatc cacctggcgg gcagcggcgc cggcgtgcac 2340 ggccagggct acgcgcacgg ggagaaggcc gtgaacaagc aagattggtt cttctcgtgc 2400 cacttctggt tcgaccccgt gatgcccggg tccctgggca tcgagtcgat gttccagctc 2460 gtcgaggcgt ggtgcgtgaa gcagggactc gcggcgcggc acggcatcgc tcacccagtg 2520 ttcgcgcacg cgcccggggc cacgagctgg aagtaccgcg ggcagctaac ccccaagaac 2580 gaccgcatgg acagcgaggt gcacatcaag tcggtggcgg ccttctcctc ctgggtcgac 2640 gtcgtcgcgg acgggttcct cttcgtcgac ggcctccgcg tctactcggc agacaacctc 2700 cgcgtccgca tccagaccgg cgccggccac gttgaagagc aagaggttgc tgccaaggcc 2760 acaaccaaga acagcagtat tgctgatgtg gacgtggcgg acctgcaagc gctcaagcag 2820 gcgttgctga cgctggagcg accgctgcag ctggacgcgg Ggagcgaggt gcccgcctgc 2880 gcggtgagcg acctgggcga taggggcttc atggagacgt acggggtggt ggcgccgctg 2940 tacagcgggg cgatggccaa gggcatcgcg tcggcggacc tggtgatcgc gatgggccag 3000

24 20103873424 201038734

cgcaagatgc tggggtcgtt tggcgcgggc gggctcccga tgcacgtcgt gcgcgcgggg attgagaaga tccaggcagc gctgccagcg gggccatacg cggtcaacct gattcactcg ccttttgacg ccaacctgga gaagggcaac gtggacctct tcctggagaa gggcgtgcgc gtcgtggagg cgtcggcctt ggcctctctc gcgacgcgcg gtcagccgca ccgagctggc aagcttgtgg cgtccggcga atggcggacg acatcgccgt cacgtcatcc tgcccctcat cctgcgcgac accgcgtgcg ctgggcgcct tccacatggg cggcaggccg ggacatgcga atcacgatgg cgccggcggc aagggcacga tgtttccctc tcgttcgagg cgatgccggc tcactcgccg aggtgtgggc gagaagatcc gcaaggcgga tggtacctcg ggctcagctc taccagatct ggtgcggccc ctcgacgtgg ccgtctcggg tcgggcgcag cctacctcca gacaccgagg acgacctctt catggagctc acgccccagg cggcggctcc gtgcgcacgg cgagatgttt atccggcccg gatcaccccc gagcaggcgg cgaggccgat tcgggcgggc cctcagcctg cgcaaccgcc cgtcggcgcc gggggcggca cgccgcgttt gtggtgacgg caatgtgcgg cggcagctgt ggacatgttc gagcagggcg gcgcgccaag aagctgttcg ggacgagctg gcgcgcgtcg cgagaccaag gacttctaca gaacgaggac cccaagctca gttctgggcc aacaacggca tgccatcggc gccttcaacg cgagttcccc gacgtcgtgc gcgcctcctc tccgtcaagc cacctaccgc cccgaccacg tggtgcgcta ccgcgcgacg cccacaagat catcggcaag cgccgcaagc cattctcgac cgctggcgct cgaggtgccc acaccgacaa ccgccccatc tccagcgcga gctcaagtac tcgggtgccc gcaagcggct gcacggtcaa ccagctgagc cgcgcgcgac gtactcggac tcgagctgca ggtgctcaag agctgtttca caagtacgac agaagcgcat cttcagcaag tcacgcggct caacaacccg agatgtcact ctgcttccgc tcgcggaccg cacgatggac acttcatcgc cgactcgtac agatcaacct gcagatcctg tcgcaccgcg gatcgacgtc cactctaa 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4308 <210> 6 <211〉 1435 Ο <212> PRT <213> 裂殖壺菌物種 <400> 6cgcaagatgc tggggtcgtt tggcgcgggc gggctcccga tgcacgtcgt gcgcgcgggg attgagaaga tccaggcagc gctgccagcg gggccatacg cggtcaacct gattcactcg ccttttgacg ccaacctgga gaagggcaac gtggacctct tcctggagaa gggcgtgcgc gtcgtggagg cgtcggcctt ggcctctctc gcgacgcgcg gtcagccgca ccgagctggc aagcttgtgg cgtccggcga atggcggacg acatcgccgt cacgtcatcc tgcccctcat cctgcgcgac accgcgtgcg ctgggcgcct tccacatggg cggcaggccg ggacatgcga atcacgatgg cgccggcggc aagggcacga tgtttccctc tcgttcgagg cgatgccggc tcactcgccg aggtgtgggc gagaagatcc gcaaggcgga tggtacctcg ggctcagctc taccagatct ggtgcggccc ctcgacgtgg ccgtctcggg tcgggcgcag cctacctcca gacaccgagg acgacctctt catggagctc acgccccagg cggcggctcc gtgcgcacgg cgagatgttt atccggcccg gatcaccccc gagcaggcgg cgaggccgat tcgggcgggc cctcagcctg cgcaaccgcc cgtcggcgcc gggggcggca cgccgcgttt gtggtgacgg caatgtgcgg cggcagctgt ggacatgttc gagcagggcg gcgcgccaag aagctgttcg ggacgagctg gcgcgcgtcg cgagaccaag gacttctaca gaacgaggac cccaagctca gttctgggcc aacaacggca tgccatcggc gccttcaacg cgagttcccc gacgtcgtgc gcgcctcctc tccgtcaagc cacctaccgc cccgaccacg tggtgcgcta ccgcgcgacg cccacaagat catcggcaag cgccgcaagc cattctcgac cgctggcgct cgaggtgccc acaccgacaa ccgccccatc tccagcgcga gctcaagtac tcgggtgccc gcaagcggct gcacggtcaa ccagctgagc cgcgcgcgac gtactcggac tcgagctgca ggtgctcaag agctgtttca caagtacgac agaagcgcat cttcagcaag tcacgcggct caacaacccg agatgtcact ctgcttccgc tcgcggaccg cacgatggac acttcatcgc cgactcgtac agatcaacct gcagatcctg tcgcaccgcg gatcgacgtc cactctaa 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4308 <210> 6 <211> 1435 Ο <212> PRT <213> Schizochytrium species <400>

Met Thr Ser Ser Lys Lys Thr Pro Val Trp Glu Met Ser Lys Glu Glu 15 10 15Met Thr Ser Ser Lys Lys Thr Pro Val Trp Glu Met Ser Lys Glu Glu 15 10 15

Leu Leu Asp Gly Lys 20Leu Leu Asp Gly Lys 20

Thr Val Val Phe Asp Tyr Asn Glu Leu 25 30Thr Val Val Phe Asp Tyr Asn Glu Leu 25 30

Leu GluLeu Glu

Phe Asp liePhe Asp lie

Phe Ala Glu Gly Asp Val Gly Gin Val Phe Gly Pro Glu 35 40 45 lie Asp Lys Tyr Arg Arg Arg Val Arg Leu Pro Ala Arg Glu Tyr Leu 50 55 60Phe Ala Glu Gly Asp Val Gly Gin Val Phe Gly Pro Glu 35 40 45 lie Asp Lys Tyr Arg Arg Arg Val Arg Leu Pro Ala Arg Glu Tyr Leu 50 55 60

Leu Val Ser Arg Val Thr Leu Met Asp Ala Glu Val Asn Asn Phe Arg 65 70 75 80Leu Val Ser Arg Val Thr Leu Met Asp Ala Glu Val Asn Asn Phe Arg 65 70 75 80

Val Gly Ser Arg Met Val Thr Glu Tyr Asp Val Pro Val Asn Gly Glu 85 90 95 25 201038734Val Gly Ser Arg Met Val Thr Glu Tyr Asp Val Pro Val Asn Gly Glu 85 90 95 25 201038734

Leu Ser Glu Gly Gly Asp Val Pro Trp Ala Val Leu Val Glu Ser Gly 100 105 110Leu Ser Glu Gly Gly Asp Val Pro Trp Ala Val Leu Val Glu Ser Gly 100 105 110

Gin Cys Asp Leu Met Leu lie Ser Tyr Met Gly lie Asp Phe Gin Cys 115 120 125Gin Cys Asp Leu Met Leu lie Ser Tyr Met Gly lie Asp Phe Gin Cys 115 120 125

Lys Gly Asp Arg Val Tyr Arg Leu Leu Asn Thr Ser Leu Thr Phe Phe 130 135 140Lys Gly Asp Arg Val Tyr Arg Leu Leu Asn Thr Ser Leu Thr Phe Phe 130 135 140

Gly Val Ala His Glu Gly Glu Thr Leu Val Tyr Asp lie Arg Val Thr 145 150 155 160Gly Val Ala His Glu Gly Glu Thr Leu Val Tyr Asp lie Arg Val Thr 145 150 155 160

Gly Phe Ala Lys Gly Ala Gly Gly Glu lie Ser Met Phe Phe Phe Glu 165 170 175Gly Phe Ala Lys Gly Ala Gly Gly Glu lie Ser Met Phe Phe Phe Glu 165 170 175

Tyr Asp Cys Phe Val Asp Gly Arg Leu Leu lie Glu Met Arg Asp Gly 180 185 190Tyr Asp Cys Phe Val Asp Gly Arg Leu Leu lie Glu Met Arg Asp Gly 180 185 190

Cys Ala Gly Phe Phe Thr Asp Ala Glu Leu Ala Ala Gly Lys Gly Val 195 200 205Cys Ala Gly Phe Phe Thr Asp Ala Glu Leu Ala Ala Gly Lys Gly Val 195 200 205

Leu Lys Thr Lys Ala Glu Leu Ala Ala Arg Ala Gin lie Gin Lys Gin 210 215 220Leu Lys Thr Lys Ala Glu Leu Ala Ala Arg Ala Gin lie Gin Lys Gin 210 215 220

Asp 工le Ala Pro Phe Ala Pro Ala Pro Cys Ser His Lys Thr Ser Leu 225 230 235 240Asp work le Ala Pro Phe Ala Pro Ala Pro Cys Ser His Lys Thr Ser Leu 225 230 235 240

Asp Ala Arg Glu Met Arg Leu Leu Val Asp Arg Gin Trp Ala Arg Val 245 250 255Asp Ala Arg Glu Met Arg Leu Leu Val Asp Arg Gin Trp Ala Arg Val 245 250 255

Phe Gly Ser Gly Met Ala Gly lie Asp Tyr Lys Leu Cys Ala Arg Lys 260 265 270Phe Gly Ser Gly Met Ala Gly lie Asp Tyr Lys Leu Cys Ala Arg Lys 260 265 270

Met Leu Met 工le Asp Arg Val Thr His Leu Asp Pro Arg Gly Gly Ala 275 280 285 1Met Leu Met work le Asp Arg Val Thr His Leu Asp Pro Arg Gly Gly Ala 275 280 285 1

His Gly Leu Gly Leu Leu lie Gly Glu Lys Val Leu Glu Arg Asp His 290 295 300His Gly Leu Gly Leu Leu lie Gly Glu Lys Val Leu Glu Arg Asp His 290 295 300

Trp Tyr Phe Pro Cys His Phe Val Arg Asp Glu Val Met Ala Gly Ser 305 310 315 320Trp Tyr Phe Pro Cys His Phe Val Arg Asp Glu Val Met Ala Gly Ser 305 310 315 320

Leu Val Ser Asp Gly Cys Ser Gin Leu Leu Lys Val Tyr Met Leu Trp 325 330 335Leu Val Ser Asp Gly Cys Ser Gin Leu Leu Lys Val Tyr Met Leu Trp 325 330 335

Leu Gly Leu His Thr Thr Val Gly Ala Phe Asp Phe Arg Pro Val Ser 340 345 350Leu Gly Leu His Thr Thr Val Gly Ala Phe Asp Phe Arg Pro Val Ser 340 345 350

Gly His Ala Asn Lys Val Arg Cys Arg Gly Gin lie Ser Pro His Lys 355 360 365Gly His Ala Asn Lys Val Arg Cys Arg Gly Gin lie Ser Pro His Lys 355 360 365

Gly Lys Leu Val Tyr Val Met Glu 工le Lys Glu Met Gly Phe Asp Ala 370 375 380Gly Lys Leu Val Tyr Val Met Glu l Lys Glu Met Gly Phe Asp Ala 370 375 380

Lys Thr Gly Asp Pro Phe Ala lie Ala Asp Val Asp lie lie Asp Val 26 201038734 385 390 395 400Lys Thr Gly Asp Pro Phe Ala lie Ala Asp Val Asp lie lie Asp Val 26 201038734 385 390 395 400

Asn Phe Glu Glu Gly Gin Ala Phe Ala Gly Val Glu Asp Leu His Ser 405 410 415Asn Phe Glu Glu Gly Gin Ala Phe Ala Gly Val Glu Asp Leu His Ser 405 410 415

Tyr Gly Gin Gly Asp Leu Arg Lys Lys lie Val Val Asp Phe Lys Gly 420 425 430 工le Ala Leu Ser Leu Gin Lys Arg Lys Glu Gin Gin Lys Glu Ser Met 435 440 445Tyr Gly Gin Gly Asp Leu Arg Lys Lys lie Val Val Asp Phe Lys Gly 420 425 430 work le Ala Leu Ser Leu Gin Lys Arg Lys Glu Gin Gin Lys Glu Ser Met 435 440 445

Thr Val Thr Thr Thr Thr Thr Thr Thr Ser Arg Val 工le Ala Pro Pro 450 455 460Thr Val Thr Thr Thr Thr Thr Thr Thr Thr Ser Arg Val A Le Pro Pro 450 455 460

Ser Gly Cys Leu Lys Gly Asp Pro Thr Ala Pro Thr Ser Val Thr Trp 465 470 475 480Ser Gly Cys Leu Lys Gly Asp Pro Thr Ala Pro Thr Ser Val Thr Trp 465 470 475 480

His Pro Met Ala Glu Gly Asn Gly Gly Pro Gly Pro Thr Pro Ser Phe 485 490 495His Pro Met Ala Glu Gly Asn Gly Gly Pro Gly Pro Thr Pro Ser Phe 485 490 495

Ser Pro Ser Ala Tyr Pro Pro Arg Ala Val Cys Phe Ser Pro Phe Pro 500 505 510Ser Pro Ser Ala Tyr Pro Pro Arg Ala Val Cys Phe Ser Pro Phe Pro 500 505 510

Asn Asn Pro Leu Asp Asn Asp His Thr Pro Gly Gin Met Pro Leu Thr 515 520 525Asn Asn Pro Leu Asp Asn Asp His Thr Pro Gly Gin Met Pro Leu Thr 515 520 525

Trp Phe Asn Met Ser Glu Phe Met Cys Gly Lys Val Ser Asn Cys Leu 530 535 540Trp Phe Asn Met Ser Glu Phe Met Cys Gly Lys Val Ser Asn Cys Leu 530 535 540

Gly Pro Glu Phe Ala Arg Phe Asp Ala Ser Lys Thr Ser Arg Ser Pro 545 550 555 560Gly Pro Glu Phe Ala Arg Phe Asp Ala Ser Lys Thr Ser Arg Ser Pro 545 550 555 560

Ala Phe Asp Leu Ala Leu Val Thr Arg Val Thr Ser Val Ala Asp Met 565 570 575Ala Phe Asp Leu Ala Leu Val Thr Arg Val Thr Ser Val Ala Asp Met 565 570 575

Glu His Gly Pro Phe Tyr Asn Val Asp Val Asn Pro Gly Gin Gly Thr 580 585 590Glu His Gly Pro Phe Tyr Asn Val Asp Val Asn Pro Gly Gin Gly Thr 580 585 590

Met Val Gly Glu Phe Asp Cys Pro Ala Asp Ala Trp Phe Phe Gly Ala 595 600 605Met Val Gly Glu Phe Asp Cys Pro Ala Asp Ala Trp Phe Phe Gly Ala 595 600 605

Ser Ser Arg Asp Asp His Met Pro Tyr Ser He Leu Met Glu He Ala 610 615 620Ser Ser Arg Asp Asp His Met Pro Tyr Ser He Leu Met Glu He Ala 610 615 620

Leu Gin Thr Ser Gly Val Leu Thr Ser Val Leu Lys Ala Pro Leu Thr 625 630 635 640Leu Gin Thr Ser Gly Val Leu Thr Ser Val Leu Lys Ala Pro Leu Thr 625 630 635 640

Met Asp Lys Asp Asp lie Leu Phe Arg Asn Leu Asp Ala Asp Ala Glu 645 650 655Met Asp Lys Asp Asp lie Leu Phe Arg Asn Leu Asp Ala Asp Ala Glu 645 650 655

Leu Val Gly Asp Ala Met Pro Asp Val Arg Gly Lys Thr lie Arg Asn 660 665 670Leu Val Gly Asp Ala Met Pro Asp Val Arg Gly Lys Thr lie Arg Asn 660 665 670

Phe Thr Lys Cys Thr Gly Tyr Ser Met Leu Gly Lys Met Gly He His 675 680 685 27 201038734Phe Thr Lys Cys Thr Gly Tyr Ser Met Leu Gly Lys Met Gly He His 675 680 685 27 201038734

Arg Phe Thr Phe Glu Leu Ser Val Asp Gly Ala Val Phe Tyr Lys Gly 690 695 700Arg Phe Thr Phe Glu Leu Ser Val Asp Gly Ala Val Phe Tyr Lys Gly 690 695 700

Ser Thr Ser Phe Gly Trp Phe Val Pro Glu Val Phe Glu Ser Gin Thr 705 710 715 720Ser Thr Ser Phe Gly Trp Phe Val Pro Glu Val Phe Glu Ser Gin Thr 705 710 715 720

Gly Leu Asp Asn Gly Lys Pro Arg Leu Pro Trp Tyr Arg Glu Asn Asn 725 730 735Gly Leu Asp Asn Gly Lys Pro Arg Leu Pro Trp Tyr Arg Glu Asn Asn 725 730 735

Val Ala Val Asp Thr Leu Ser Ala Pro Ala Ser Ala Ser Ser Ala Gin 740 745 750Val Ala Val Asp Thr Leu Ser Ala Pro Ala Ser Ala Ser Ser Ala Gin 740 745 750

Gly Gin Leu Gin Leu Gin Arg Arg Gly Ser Gin Ala Gin Phe Leu Asp 755 760 765Gly Gin Leu Gin Leu Gin Arg Arg Gly Ser Gin Ala Gin Phe Leu Asp 755 760 765

Thr lie His Leu Ala Gly Ser Gly Ala Gly Val His Gly Gin Gly Tyr 770 775 780Thr lie His Leu Ala Gly Ser Gly Ala Gly Val His Gly Gin Gly Tyr 770 775 780

Ala His Gly Glu Lys Ala Val Asn Lys Gin Asp Trp Phe Phe Ser Cys 785 790 795 800Ala His Gly Glu Lys Ala Val Asn Lys Gin Asp Trp Phe Phe Ser Cys 785 790 795 800

His Phe Trp Phe Asp Pro Val Met Pro Gly Ser Leu Gly lie Glu Ser 805 810 815His Phe Trp Phe Asp Pro Val Met Pro Gly Ser Leu Gly lie Glu Ser 805 810 815

Met Phe Gin Leu Val Glu Ala Trp Cys Val Lys Gin Gly Leu Ala Ala 820 825 830Met Phe Gin Leu Val Glu Ala Trp Cys Val Lys Gin Gly Leu Ala Ala 820 825 830

Arg His Gly lie Ala His Pro Val Phe Ala His Ala Pro Gly Ala Thr 835 840 845Arg His Gly lie Ala His Pro Val Phe Ala His Ala Pro Gly Ala Thr 835 840 845

Ser Trp Lys Tyr Arg Gly Gin Leu Thr Pro Lys Asn Asp Arg Met Asp 850 855 860Ser Trp Lys Tyr Arg Gly Gin Leu Thr Pro Lys Asn Asp Arg Met Asp 850 855 860

Ser Glu Val His lie Lys Ser Val Ala Ala Phe Ser Ser Trp Val Asp 865 870 875 880Ser Glu Val His lie Lys Ser Val Ala Ala Phe Ser Ser Trp Val Asp 865 870 875 880

Val Val Ala Asp Gly Phe Leu Phe Val Asp Gly Leu Arg Val Tyr Ser 885 890 895Val Val Ala Asp Gly Phe Leu Phe Val Asp Gly Leu Arg Val Tyr Ser 885 890 895

Ala Asp Asn Leu Arg Val Arg lie Gin Thr Gly Ala Gly His Val Glu 900 905 910Ala Asp Asn Leu Arg Val Arg lie Gin Thr Gly Ala Gly His Val Glu 900 905 910

Glu Gin Glu Val Ala Ala Lys Ala Thr Thr Lys Asn Ser Ser lie Ala 915 920 925Glu Gin Glu Val Ala Ala Lys Ala Thr Thr Lys Asn Ser Ser lie Ala 915 920 925

Asp Val Asp Val Ala Asp Leu Gin Ala Leu Lys Gin Ala Leu Leu Thr 930 935 940Asp Val Asp Val Ala Asp Leu Gin Ala Leu Lys Gin Ala Leu Leu Thr 930 935 940

Leu Glu Arg Pro Leu Gin Leu Asp Ala Gly Ser Glu Val Pro Ala Cvs 945 950 955 960Leu Glu Arg Pro Leu Gin Leu Asp Ala Gly Ser Glu Val Pro Ala Cvs 945 950 955 960

Ala Val Ser Asp Leu Gly Asp Arg Gly Phe Met Glu Thr Tyr Gly Val 965 970 975Ala Val Ser Asp Leu Gly Asp Arg Gly Phe Met Glu Thr Tyr Gly Val 965 970 975

Val Ala Pro Leu Tyr Ser Gly Ala Met Ala Lys Gly He Ala Ser Ala 980 985 990 28 201038734Val Ala Pro Leu Tyr Ser Gly Ala Met Ala Lys Gly He Ala Ser Ala 980 985 990 28 201038734

Asp Leu Val lie Ala Met Gly Gin Arg Lys Met Leu Gly Ser Phe Gly 995 1000 1005Asp Leu Val lie Ala Met Gly Gin Arg Lys Met Leu Gly Ser Phe Gly 995 1000 1005

Ala Gly Gly Leu Pro Met His Val Val 1010 1015 Arg Ala Gly lie Glu Lys 1020 lie Gin Ala Ala Leu Pro Ala Gly Pro 1025 1030 Tyr Ala Val Asn Leu lie 1035 His Ser Pro Phe Asp Ala Asn Leu Glu 1040 1045 Lys Gly Asn Val Asp Leu 1050 Phe Leu Glu Lys Gly Val Arg Val Val 1055 1060 Glu Ala Ser Ala Phe Met 1065 Glu Leu Thr Pro Gin Val Val Arg Tyr 1070 1075 Arg Ala Thr Gly Leu Ser 1080 〇 Arg Asp Ala Arg Gly Gly Ser Val Arg 1085 1090 Thr Ala His Lys lie lie 1095 Gly Lys Val Ser Arg Thr Glu Leu Ala 1100 1105 Glu Met Phe lie Arg Pro 1110 Ala Pro Gin Ala lie Leu Asp Lys Leu * 1115 1120 Val Ala Ser Gly Glu lie 1125 • Thr Pro Glu Gin Ala Ala Leu Ala Leu ^ 1130 1135 Glu Val Pro Met Ala Asp 1140 Asp lie Ala Val Glu Ala Asp Ser Gly 1145 1150 Gly His Thr Asp Asn Arg 1155 Pro lie His Val lie Leu Pro Leu lie 1160 1165 Leu Ser Leu Arg Asn Arg 1170 Leu Gin Arg Glu Leu Lys Tyr Pro Ala 1175 1180 Arg His Arg Val Arg Val 1185 Gly Ala Gly Gly Gly lie Gly Cys Pro 1190 1195 Gin Ala Ala Leu Gly Ala 1200 Phe His Met Gly Ala Ala Phe Val Val 1205 1210 Thr Gly Thr Val Asn Gin 1215 Leu Ser Arg Gin Ala Gly Thr Cys Asp 1220 1225 Asn Val Arg Arg Gin Leu 1230 Ser Arg Ala Thr Tyr Ser Asp 工le Thr 1235 1240 Met Ala Pro Ala Ala Asp 1245Ala Gly Gly Leu Pro Met His Val Val 1010 1015 Arg Ala Gly lie Glu Lys 1020 lie Gin Ala Ala Leu Pro Ala Gly Pro 1025 1030 Tyr Ala Val Asn Leu lie 1035 His Ser Pro Phe Asp Ala Asn Leu Glu 1040 1045 Lys Gly Asn Val Asp Leu 1050 Phe Leu Glu Lys Gly Val Arg Val Val 1055 1060 Glu Ala Ser Ala Phe Met 1065 Glu Leu Thr Pro Gin Val Val Arg Tyr 1070 1075 Arg Ala Thr Gly Leu Ser 1080 〇Arg Asp Ala Arg Gly Gly Ser Val Arg 1085 1090 Thr Ala His Lys lie lie 1095 Gly Lys Val Ser Arg Thr Glu Leu Ala 1100 1105 Glu Met Phe lie Arg Pro 1110 Ala Pro Gin Ala lie Leu Asp Lys Leu * 1115 1120 Val Ala Ser Gly Glu lie 1125 • Thr Pro Glu Gin Ala Ala Leu Ala Leu ^ 1130 1135 Glu Val Pro Met Ala Asp 1140 Asp lie Ala Val Glu Ala Asp Ser Gly 1145 1150 Gly His Thr Asp Asn Arg 1155 Pro lie His Val lie Leu Pro Leu lie 1160 1165 Leu Ser Leu Arg Asn Arg 1170 Leu Gin Arg Glu Leu Lys Tyr Pro Ala 1175 1180 Arg His Arg Val Arg Val 1185 Gly Ala Gly Gly Gly Gly Cys Pro 1190 1195 Gin Ala Ala Leu Gly Ala 1200 Phe His Met Gly Ala Ala Phe Val Val 1205 1210 Thr Gly Thr Val Asn Gin 1215 Leu Ser Arg Gin Ala Gly Thr Cys Asp 1220 1225 Asn Val Arg Arg Gin Leu 1230 Ser Arg Ala Thr Tyr Ser Asp worker le Thr 1235 1240 Met Ala Pro Ala Ala Asp 1245

Met Phe Glu Gin Gly Val Glu Leu Gin Val Leu Lys Lys Gly Thr 1250 1255 1260Met Phe Glu Gin Gly Val Glu Leu Gin Val Leu Lys Lys Gly Thr 1250 1255 1260

Met Phe Pro Ser Arg Ala Lys Lys Leu Phe Glu Leu Phe His Lys 1265 1270 1275 29 201038734Met Phe Pro Ser Arg Ala Lys Lys Leu Phe Glu Leu Phe His Lys 1265 1270 1275 29 201038734

Tyr Asp Ser Phe Glu Ala Met Pro Ala Asp Glu Leu Ala Arg Val 1280 1285 1290Tyr Asp Ser Phe Glu Ala Met Pro Ala Asp Glu Leu Ala Arg Val 1280 1285 1290

Glu Lys Arg lie Phe Ser Lys Ser Leu Ala Glu Val Trp Ala Glu 1295 1300 1305Glu Lys Arg lie Phe Ser Lys Ser Leu Ala Glu Val Trp Ala Glu 1295 1300 1305

Thr Lys Asp Phe Tyr lie Thr Arg Leu Asn Asn Pro Glu Lys lie 1310 1315 1320Thr Lys Asp Phe Tyr lie Thr Arg Leu Asn Asn Pro Glu Lys lie 1310 1315 1320

Arg Lys Ala Glu Asn Glu Asp Pro Lys Leu Lys Met Ser Leu Cys 1325 1330 1335Arg Lys Ala Glu Asn Glu Asp Pro Lys Leu Lys Met Ser Leu Cys 1325 1330 1335

Phe Arg Trp Tyr Leu Gly Leu Ser Ser Phe Trp Ala Asn Asn Gly 1340 1345 1350Phe Arg Trp Tyr Leu Gly Leu Ser Ser Phe Trp Ala Asn Asn Gly 1340 1345 1350

lie Ala Asp Arg Thr Met Asp Tyr Gin lie Trp Cys Gly Pro Ala 1355 1360 1365 工le Gly Ala Phe Asn Asp Phe 工le Ala Asp Ser Tyr Leu Asp Val 1370 1375 1380Lie Ala Asp Arg Thr Met Asp Tyr Gin lie Trp Cys Gly Pro Ala 1355 1360 1365 work le Gly Ala Phe Asn Asp Phe work le Ala Asp Ser Tyr Leu Asp Val 1370 1375 1380

Ala Val Ser Gly Glu Phe Pro Asp Val Val Gin lie Asn Leu Gin 1385 1390 1395 工le Leu Ser Gly Ala Ala Tyr Leu Gin Arg Leu Leu Ser Val Lys 1400 1405 1410Ala Val Ser Gly Glu Phe Pro Asp Val Val Gin lie Asn Leu Gin 1385 1390 1395 Le Leu Ser Gly Ala Ala Tyr Leu Gin Arg Leu Leu Ser Val Lys 1400 1405 1410

Leu Ala Pro Arg lie Asp Val Asp Thr Glu Asp Asp Leu Phe Thr 1415 1420 1425Leu Ala Pro Arg lie Asp Val Asp Thr Glu Asp Asp Leu Phe Thr 1415 1420 1425

Tyr Arg Pro Asp His Ala Leu 1430 1435 <21〇> 7 <211> 1395Tyr Arg Pro Asp His Ala Leu 1430 1435 <21〇> 7 <211> 1395

<212> DNA <213〉 裂殖壺菌物種 <400> 7 actcgcatcg cgatcgtggg gatgtcggcg atcctgccga gcggggagaa cgtgcgcgag 60 agctgggagg cgatccgcga tgggctggat tgcctgagcg atctgccggc ggaccgcgtg 120 gacgtgacgg cctactacaa cccggagaag acgaccaagg acaagatcta ctgcaagcgc 180 ggcgggttca tcccggagta cgacttcgac gcgcgtgagt tcgggctcaa catgttccag 240 atggaggact cggacgccaa ccagacgatc tcgctgctca aggtgaagga ggcgctgacg 300 gacgccaaca tcccggcgtt ctcgagcggt aagaagaaca tcggctgcgt gctgggcatc 360 ggcggcggcc agaaggcgag ccacgagttc tactcgcggc tcaactacgt ggtcgtggac 420 aaggtgctgc gcaagatggg cctgccggag gaagacgtgg cggcggcggt ggacaagtac 480 aaggcgagtt tccccgagtg gcgcctcgac tctttccccg ggttcctggg caacgtcacg 540 gcggggcgct gctgcaatac cttcaacatg gagggcatga actgcgtcgt ggacgcggcc 600 tgcgcgtcgt cgctgatcgc ggtcaaagtg gcgatcgagg agctgctcta cggcgactgc 660 gatgcgatga tcgcgggtgc cacctgcacg gacaactcga tcgggatgta catggccttc 720 30 780 201038734cggacgccaa ccagacgatc 7 actcgcatcg cgatcgtggg gatgtcggcg atcctgccga gcggggagaa cgtgcgcgag 60 agctgggagg cgatccgcga tgggctggat tgcctgagcg atctgccggc ggaccgcgtg 120 gacgtgacgg cctactacaa cccggagaag acgaccaagg acaagatcta ctgcaagcgc 180 ggcgggttca tcccggagta cgacttcgac gcgcgtgagt tcgggctcaa catgttccag 240 atggaggact; < 212 > DNA < 213> Schizochytrium species < 400 & gt tcgctgctca aggtgaagga ggcgctgacg 300 gacgccaaca tcccggcgtt ctcgagcggt aagaagaaca tcggctgcgt gctgggcatc 360 ggcggcggcc agaaggcgag ccacgagttc tactcgcggc tcaactacgt ggtcgtggac 420 aaggtgctgc gcaagatggg cctgccggag gaagacgtgg cggcggcggt ggacaagtac 480 aaggcgagtt tccccgagtg gcgcctcgac tctttccccg ggttcctggg caacgtcacg 540 gcggggcgct gctgcaatac cttcaacatg gagggcatga actgcgtcgt ggacgcggcc 600 tgcgcgtcgt cgctgatcgc ggtcaaagtg gcgatcgagg agctgctcta cggcgactgc 660 gatgcgatga tcgcgggtgc cacctgcacg gacaactcga Tcgggatgta catggccttc 720 30 780 201038734

840 900 960 1020 1080 1140 1200 1260 1320 1380 1395 tccaagacgc ccgtgttttc cacggacccg agcgtcaagg cgtacgacgc cgccaccaaa ggcatgctca tcggcgaggg ctcggcgatg ctcgtgctga agcgctacgc ggacgccgtg cgcgacggcg acaccgtgca cgccgtcatc aaggggtgcg cgtcctcgag cgacggcaag gcggcgggca tctacacgcc gacaatctcg ggccaggagg aggccctgcg ccgcgcctac gcccgcgcca atgtcgaccc ggccactgtg acgctggtgg agggccacgg cacgggtacg ccggtgggcg acaagatcga gctgacggcg ctgagcaacc tcttctccaa ggcgttttct gccaacggtg gcggcgcgga ggaagcagag caggtggcgg tgggcagcat caagtcgcag atcgggcacc tcaaggcggt ggccgggctg gccgggctgg tcaaggtggt gctggcgctc aagcacaaga cgctgccgca gacgatcaac gtcgacaagc cgccgtcgct ggtggacggg accccgatcc agcagtcgcc gctgtacgtc aacacgatga accgcccctg gttcacgccc gtaggggtgc cgcgccgcgc cggcgtgtcg tcgtttgggt ttggcggtgc caactaccac gccgtgctgg aggag <21〇> 8 <211> 465 <212> PRT <213〉 裂殖壺菌物種 <400> 8840 900 960 1020 1080 1140 1200 1260 1320 1380 1395 tccaagacgc ccgtgttttc cacggacccg agcgtcaagg cgtacgacgc cgccaccaaa ggcatgctca tcggcgaggg ctcggcgatg ctcgtgctga agcgctacgc ggacgccgtg cgcgacggcg acaccgtgca cgccgtcatc aaggggtgcg cgtcctcgag cgacggcaag gcggcgggca tctacacgcc gacaatctcg ggccaggagg aggccctgcg ccgcgcctac gcccgcgcca atgtcgaccc ggccactgtg acgctggtgg agggccacgg cacgggtacg ccggtgggcg acaagatcga gctgacggcg ctgagcaacc tcttctccaa ggcgttttct gccaacggtg gcggcgcgga ggaagcagag caggtggcgg tgggcagcat caagtcgcag atcgggcacc tcaaggcggt ggccgggctg gccgggctgg tcaaggtggt gctggcgctc aagcacaaga cgctgccgca gacgatcaac gtcgacaagc cgccgtcgct ggtggacggg accccgatcc agcagtcgcc gctgtacgtc aacacgatga accgcccctg gttcacgccc gtaggggtgc cgcgccgcgc cggcgtgtcg tcgtttgggt ttggcggtgc caactaccac gccgtgctgg aggag < 21〇 > 8 < 211 > 465 < 212 > PRT < 213> Schizochytrium species <400> 8

Thr Arg lie Ala lie Val Gly Met Ser Ala lie Leu Pro Ser Gly Glu 15 10 15Thr Arg lie Ala lie Val Gly Met Ser Ala lie Leu Pro Ser Gly Glu 15 10 15

Asn Val Arg Glu Ser Trp Glu Ala lie Arg Asp Gly Leu Asp Cys Leu 20 25 30Asn Val Arg Glu Ser Trp Glu Ala lie Arg Asp Gly Leu Asp Cys Leu 20 25 30

Ser Asp Leu Pro Ala Asp Arg Val Asp Val Thr Ala Tyr Tyr Asn Pro 35 40 45Ser Asp Leu Pro Ala Asp Arg Val Asp Val Thr Ala Tyr Tyr Asn Pro 35 40 45

Glu Lys Thr Thr Lys Asp Lys He Tyr Cys Lys Arg Gly Gly Phe lie 50 55 60Glu Lys Thr Thr Lys Asp Lys He Tyr Cys Lys Arg Gly Gly Phe lie 50 55 60

Pro Glu Tyr Asp Phe Asp Ala Arg Glu Phe Gly Leu Asn Met Phe Gin 65 70 75 80Pro Glu Tyr Asp Phe Asp Ala Arg Glu Phe Gly Leu Asn Met Phe Gin 65 70 75 80

Met Glu Asp Ser Asp Ala Asn Gin Thr lie Ser Leu Leu Lys Val Lys 85 90 95Met Glu Asp Ser Asp Ala Asn Gin Thr lie Ser Leu Leu Lys Val Lys 85 90 95

Glu Ala Leu Thr Asp Ala Asn lie Pro Ala Phe Ser Ser Gly Lys Lys 100 105 110Glu Ala Leu Thr Asp Ala Asn lie Pro Ala Phe Ser Ser Gly Lys Lys 100 105 110

Asn lie Gly Cys Val Leu Gly lie Gly Gly Gly Gin Lys Ala Ser His 115 120 125Asn lie Gly Cys Val Leu Gly lie Gly Gly Gly Gin Lys Ala Ser His 115 120 125

Glu Phe Tyr Ser Arg Leu Asn Tyr Val Val Val Asp Lys Val Leu Arq 130 135 140Glu Phe Tyr Ser Arg Leu Asn Tyr Val Val Val Asp Lys Val Leu Arq 130 135 140

Lys Met Gly Leu Pro Glu Glu Asp Val Ala Ala Ala Val Asp Lys Tyr 145 150 155 160Lys Met Gly Leu Pro Glu Glu Asp Val Ala Ala Ala Val Asp Lys Tyr 145 150 155 160

Lys Ala Ser Phe Pro Glu Trp Arg Leu Asp Ser Phe Pro Gly Phe Leu 165 170 175 31 201038734Lys Ala Ser Phe Pro Glu Trp Arg Leu Asp Ser Phe Pro Gly Phe Leu 165 170 175 31 201038734

Gly Asn Val Thr Ala Gly Arg Cys Cys Asn Thr Phe Asn Met Glu Gly 180 185 190Gly Asn Val Thr Ala Gly Arg Cys Cys Asn Thr Phe Asn Met Glu Gly 180 185 190

Met Asn Cys Val Val Asp Ala Ala Cys Ala Ser Ser Leu lie Ala Val 195 200 205Met Asn Cys Val Val Asp Ala Ala Cys Ala Ser Ser Leu lie Ala Val 195 200 205

Lys Val Ala lie Glu Glu Leu Leu Tyr Gly Asp Cys Asp Ala Met lie 210 215 220Lys Val Ala lie Glu Glu Leu Leu Tyr Gly Asp Cys Asp Ala Met lie 210 215 220

Ala Gly Ala Thr Cys Thr Asp Asn Ser lie Gly Met Tyr Met Ala Phe 225 230 235 240Ala Gly Ala Thr Cys Thr Asp Asn Ser lie Gly Met Tyr Met Ala Phe 225 230 235 240

Ser Lys Thr Pro Val Phe Ser Thr Asp Pro Ser Val Lys Ala Tyr Asp 245 250 255Ser Lys Thr Pro Val Phe Ser Thr Asp Pro Ser Val Lys Ala Tyr Asp 245 250 255

Ala Ala Thr Lys Gly Met Leu lie Gly Glu Gly Ser Ala Met Leu Val 260 265 270Ala Ala Thr Lys Gly Met Leu lie Gly Glu Gly Ser Ala Met Leu Val 260 265 270

))

Leu Lys Arg Tyr Ala Asp Ala Val Arg Asp Gly Asp Thr Val His Ala 275 280 285Leu Lys Arg Tyr Ala Asp Ala Val Arg Asp Gly Asp Thr Val His Ala 275 280 285

Val lie Lys Gly Cys Ala Ser Ser Ser Asp Gly Lys Ala Ala Gly lie 290 295 300Val lie Lys Gly Cys Ala Ser Ser Ser Asp Gly Lys Ala Ala Gly lie 290 295 300

Tyr Thr Pro Thr lie Ser Gly Gin Glu Glu Ala Leu Arg Arg Ala Tyr 305 310 315 320Tyr Thr Pro Thr lie Ser Gly Gin Glu Glu Ala Leu Arg Arg Ala Tyr 305 310 315 320

Ala Arg Ala Asn Val Asp Pro Ala Thr Val Thr Leu Val Glu Gly His 325 330 335Ala Arg Ala Asn Val Asp Pro Ala Thr Val Thr Leu Val Glu Gly His 325 330 335

Gly Thr Gly Thr Pro Val Gly Asp Lys lie Glu Leu Thr Ala Leu Ser 340 345 350Gly Thr Gly Thr Pro Val Gly Asp Lys lie Glu Leu Thr Ala Leu Ser 340 345 350

Asn Leu Phe Ser Lys Ala Phe Ser Ala Asn Gly Gly Gly Ala Glu Glu 355 360 365Asn Leu Phe Ser Lys Ala Phe Ser Ala Asn Gly Gly Gly Ala Glu Glu 355 360 365

Ala Glu Gin Val Ala Val Gly Ser lie Lys Ser Gin lie Gly His Leu 370 375 380Ala Glu Gin Val Ala Val Gly Ser lie Lys Ser Gin lie Gly His Leu 370 375 380

Lys Ala Val Ala Gly Leu Ala Gly Leu Val Lys Val Val Leu Ala Leu 385 390 395 400Lys Ala Val Ala Gly Leu Ala Gly Leu Val Lys Val Val Leu Ala Leu 385 390 395 400

Lys His Lys Thr Leu Pro Gin Thr lie Asn Val Asp Lys Pro Pro Ser 405 410 415Lys His Lys Thr Leu Pro Gin Thr lie Asn Val Asp Lys Pro Pro Ser 405 410 415

Leu Val Asp Gly Thr Pro lie Gin Gin Ser Pro Leu Tyr Val Asn Thr 420 425 430Leu Val Asp Gly Thr Pro lie Gin Gin Ser Pro Leu Tyr Val Asn Thr 420 425 430

Met Asn Arg Pro Trp Phe Thr Pro Val Gly Val Pro Arg Arg Ala Gly 435 440 445Met Asn Arg Pro Trp Phe Thr Pro Val Gly Val Pro Arg Arg Ala Gly 435 440 445

Val Ser Ser Phe Gly Phe Gly Gly Ala Asn Tyr His Ala Val Leu Glu 450 455 460Val Ser Ser Phe Gly Phe Gly Gly Ala Asn Tyr His Ala Val Leu Glu 450 455 460

Glu 32 60 60Glu 32 60 60

201038734 465 <210> 9 <211> 903 <212> DNA <213〉 裂殖壺菌物種 <400〉 9 ttctcgggcc agggcgcgca gtacacgcac atgttcagcg acgtggcgat gaactggccc ccgttccgcg agagcgtcgc cgccatggac cgcgcccagc gcgagcgctt cgggcggcct gccaagcgcg tgagcagcgt gctgtacccg cgcaagccgt acggcgacga accgcggcag gaccacaagg agatctcgca aacgcgctac tcgcagcccg caacgctcgc gtgctcggtc ggcgcctttg acatcttcaa agcggcggga ctggcgccga gctttgcggc gggccactcg ctgggcgagt ttgcggcgct ctacgcggcc gggtcgctcg atcgcgacgc cgtcttcgac ctggtctgcg cgcgcgccaa ggccatgagc gacttcacgg cccaggccag cagcagcggt ggcgccatgg cggccgtgat tggcgccaag gcggaccagc tctcgctggg tggcgcgccc gacgtgtggc tcgccaacag caactcgccc tcgcagaccg tgatcacggg aaccgccgaa gcagtggctg cggcctctga caagttgcgc tgcagcggca acttccgcgt cgtgcctctg gcctgcgagg cggccttcca ctcgccgcac atgcgcggcg cggagcagac gtttgcgtcg gcgctcgcgc aggcgcccgt gtcggcaccg gcggctgctc ggttctactc taacgtgacg gggggcgccg cggtaacctc gcccgcggac gtcaaaacga acctgggcaa gcacatgacg agccctgtgc agttcgtgca gcaggtgcga gccatgcacg cggcgggcgc gcgtgtgttt gtggagtttg ggcccaagca ggtcctgtcg cgcctcgtca aggagaccct tggcgaggcc ggc <210> 10 <211> 301 <212> PRT <213〉 裂殖壺菌物種 <400> 10201038734 465 < 210 > 9 < 211 > 903 < 212 > DNA < 213> Schizochytrium species < 400> 9 ttctcgggcc agggcgcgca gtacacgcac atgttcagcg acgtggcgat gaactggccc ccgttccgcg agagcgtcgc cgccatggac cgcgcccagc gcgagcgctt cgggcggcct gccaagcgcg tgagcagcgt gctgtacccg cgcaagccgt acggcgacga accgcggcag gaccacaagg agatctcgca aacgcgctac tcgcagcccg caacgctcgc gtgctcggtc ggcgcctttg acatcttcaa agcggcggga ctggcgccga gctttgcggc gggccactcg ctgggcgagt ttgcggcgct ctacgcggcc gggtcgctcg atcgcgacgc cgtcttcgac ctggtctgcg cgcgcgccaa ggccatgagc gacttcacgg cccaggccag cagcagcggt ggcgccatgg cggccgtgat tggcgccaag gcggaccagc tctcgctggg tggcgcgccc gacgtgtggc tcgccaacag caactcgccc tcgcagaccg tgatcacggg aaccgccgaa gcagtggctg cggcctctga caagttgcgc tgcagcggca acttccgcgt cgtgcctctg gcctgcgagg cggccttcca ctcgccgcac atgcgcggcg cggagcagac gtttgcgtcg gcgctcgcgc aggcgcccgt gtcggcaccg Gcggctgctc ggttctactc taacgtgacg gggggcgccg cggtaacctc gcccgcggac gtcaaaacga acctgggcaa gcacatgacg agccctgtgc agttcgtgca gcaggtgcga gccatg Cacg cggcgggcgc gcgtgtgttt gtggagtttg ggcccaagca ggtcctgtcg cgcctcgtca aggagaccct tggcgaggcc ggc <210> 10 <211> 301 <212> PRT <213> Schizochytrium species <400>

Phe Ser Gly Gin Gly Ala Gin Tyr Thr His Met Phe Ser Asp Val Ala 15 10 15Phe Ser Gly Gin Gly Ala Gin Tyr Thr His Met Phe Ser Asp Val Ala 15 10 15

Met Asn Trp Pro Pro Phe Arg Glu Ser Val Ala Ala Met Asp Arg Ala 20 25 30Met Asn Trp Pro Pro Phe Arg Glu Ser Val Ala Ala Met Asp Arg Ala 20 25 30

Gin Arg Glu Arg Phe Gly Arg Pro Ala Lys Arg Val Ser Ser Val Leu 35 40 45Gin Arg Glu Arg Phe Gly Arg Pro Ala Lys Arg Val Ser Ser Val Leu 35 40 45

Tyr Pro Arg Lys Pro Tyr Gly Asp Glu Pro Arg Gin Asp His Lys Glu 50 55 60 lie Ser Gin Thr Arg Tyr Ser Gin Pro Ala Thr Leu Ala Cys Ser Val 65 70 75 80Tyr Pro Arg Lys Pro Tyr Gly Asp Glu Pro Arg Gin Asp His Lys Glu 50 55 60 lie Ser Gin Thr Arg Tyr Ser Gin Pro Ala Thr Leu Ala Cys Ser Val 65 70 75 80

Gly Ala Phe Asp lie Phe Lys Ala Ala Gly Leu Ala Pro Ser Phe Ala 85 90 95Gly Ala Phe Asp lie Phe Lys Ala Ala Gly Leu Ala Pro Ser Phe Ala 85 90 95

Ala Gly His Ser Leu Gly Glu Phe Ala Ala Leu Tyr Ala Ala Gly Ser 100 105 110 120 180 240 300 360 420 480 540 600 660 720 780 840 900 903 33 201038734Ala Gly His Ser Leu Gly Glu Phe Ala Ala Leu Tyr Ala Ala Gly Ser 100 105 110 120 180 240 300 360 420 480 540 600 660 720 780 840 900 903 33 201038734

Leu Asp Arg Asp Ala Val Phe Asp Leu Val Cys Ala Arg Ala Lys Ala 115 120 125 Met Ser Asp Phe Thr Ala Gin Ala Ser Ser Ser Gly Gly Ala Met Ala 130 135 140 Ala Val lie Gly Ala Lys Ala Asp Gin Leu Ser Leu Gly Gly Ala Pro 145 150 155 160 Asp Val Trp Leu Ala Asn Ser Asn Ser Pro Ser Gin Thr Val lie Thr 165 170 175 Gly Thr Ala Glu Ala Val Ala Ala Ala Ser Asp Lys Leu Arg Cys Ser 180 185 190Leu Asp Arg Asp Ala Val Phe Asp Leu Val Cys Ala Arg Ala Lys Ala 115 120 125 Met Ser Asp Phe Thr Ala Gin Ala Ser Ser Ser Gly Gly Ala Met Ala 130 135 140 Ala Val lie Gly Ala Lys Ala Asp Gin Leu Ser Leu Gly Gly Ala Pro 145 150 155 160 Asp Val Trp Leu Ala Asn Ser Asn Ser Pro Ser Gin Thr Val lie Thr 165 170 175 Gly Thr Ala Glu Ala Val Ala Ala Ala Ser Asp Lys Leu Arg Cys Ser 180 185 190

Gly Asn Phe Arg Val Val Pro Leu Ala Cys Glu Ala Ala Phe His Ser 195 200 205Gly Asn Phe Arg Val Val Pro Leu Ala Cys Glu Ala Ala Phe His Ser 195 200 205

OO

Pro His Met Arg Gly Ala Glu Gin Thr Phe Ala 210 215 n H G a i—I A u e L a Ι Α r o e 2 s 2Pro His Met Arg Gly Ala Glu Gin Thr Phe Ala 210 215 n H G a i-I A u e L a Ι Α r o e 2 s 2

Ala Pro Val Ser Ala Pro Ala Ala Ala Arg Phe Tyr Ser Asn Val Thr 225 230 235 240Ala Pro Val Ser Ala Pro Ala Ala Ala Arg Phe Tyr Ser Asn Val Thr 225 230 235 240

Gly Gly Ala Ala Val Thr Ser Pro Ala Asp Val Lys Thr Asn Leu Gly 245 250 255 a V e 5 h6 p 2 n _—I G 1 a V 〇 r p r e s r o h 6 T 2 t e M s i H s yGly Gly Ala Ala Val Thr Ser Pro Ala Asp Val Lys Thr Asn Leu Gly 245 250 255 a V e 5 h6 p 2 n _—I G 1 a V 〇 r p r e s r o h 6 T 2 t e M s i H s y

Gin Gin Val Arg Ala Met 270Gin Gin Val Arg Ala Met 270

His Ala Ala Gly Ala Arg Val Phe Val Glu Phe Gly Pro Lys Gin Val 275 280 285His Ala Ala Gly Ala Arg Val Phe Val Glu Phe Gly Pro Lys Gin Val 275 280 285

Leu Ser Arg Leu Val Lys Glu Thr Leu Gly Glu Ala Gly 290 295 300 <210> 11 <211> 2103 <212> DNA <213> 裂殖壺菌物種 <400> 11 tccggcaaca gcaagagcac tcgtggcagt gctgatctgc aagcgctgct ggccaaggcg 60 gagactgtgg tgatggctgt gctggctgcc aagactggct acgaggccga catggttgag 120 gcggacatgg acctggaggc cgagctcggc atcgactcga tcaagcgcgt ggagatcctt 180 tccgaggtgc agggccagct gggcgtcgag gccaaggacg tggatgcgct gagccgcacg 240 cgcacggtcg gtgaggttgt ggacgccatg aaggcggaga tcgtggctgc ctctggtggt 300 agtgctcctg cggttccttc ggcgcccgct gcttctgcag ctccgactcc cgctgcttcg 360 actgcgcctt ctgctgatct gcaagcgctg ctgtccaagg cggagactgt ggtgatggct 420 gtgctggcgg ccaagactgg ctacgaggcc gacatggtcg aggcggacat ggacctggag 480 gccgagctcg gcatcgactc gatcaagcgc gtggagatcc tctcggaggt gcagggccag 540 ctgggcgtcg aggccaagga cgtggatgcg ctgagccgca cgcgcacggt cggtgaggtt 600 34 660 660 ΟLeu Ser Arg Leu Val Lys Glu Thr Leu Gly Glu Ala Gly 290 295 300 <210> 11 <211> 2103 <212> DNA <213> Schizochytrium species <400> 11 tccggcaaca gcaagagcac tcgtggcagt gctgatctgc aagcgctgct ggccaaggcg 60 gagactgtgg tgatggctgt gctggctgcc aagactggct acgaggccga catggttgag 120 gcggacatgg acctggaggc cgagctcggc atcgactcga tcaagcgcgt ggagatcctt 180 tccgaggtgc agggccagct gggcgtcgag gccaaggacg tggatgcgct gagccgcacg 240 cgcacggtcg gtgaggttgt ggacgccatg aaggcggaga tcgtggctgc ctctggtggt 300 agtgctcctg cggttccttc ggcgcccgct gcttctgcag ctccgactcc cgctgcttcg 360 actgcgcctt ctgctgatct gcaagcgctg ctgtccaagg cggagactgt ggtgatggct 420 gtgctggcgg ccaagactgg ctacgaggcc gacatggtcg aggcggacat ggacctggag 480 gccgagctcg gcatcgactc gatcaagcgc gtggagatcc tctcggaggt gcagggccag 540 ctgggcgtcg aggccaagga cgtggatgcg ctgagccgca cgcgcagggt cggtgaggtt 600 34 660 660 Ο

G 201038734 gtggatgcca tgaaggcgga aatcgtggct gcctctgctg gtagtgctcc tgctcctgct gttccttcgg cgcccgctgc ttctgcagct ccgactcccg ctgcttcgac tgcgccttct gctgatctgc aagcgctgct gtccaaggcg gagacggtgg tgatggctgt gctggcggcc aagactggct acgaggccga catggtcgag gcggacatgg acctggaggc cgagctcggc atcgactcga tcaagcgcgt ggagatcctc tcggaggtgc agggccagct gggcgtcgag gccaaggacg tggatgcgct gagccgcacg cgcacggtcg gtgaggttgt ggatgccatg aaggcggaaa tcgtggctgc ctctggtggt agtgctcctg ctcctgcggt tccttcggcg cccgctgctt ctgcagctcc gactcccgcg gctgcgacag cgccttctgc tgatctgcaa gcgctgctgg ccaaggcgga gactgtggtg atggctgtgc tggcggccaa gactggctac gaggccgaca tggtcgaggc ggacatggac ctggaggccg agctcggcat cgactcgatc aagcgcgtgg agatcctttc cgaggtgcag ggccagctgg gcgtcgaggc caaggacgta gatgcgctga gccgcacgcg cacggtcggt gaggttgtgg atgccatgaa ggcggagatc gtggctgcct ctgctggtag tgctcctgct cctgctgttc cttcggcgcc cgctgcttct gcagctccga ctcccgctgc ttcgactgcg ccttctgctg atctgcaagc gctgctgtcc aaggcggaga ctgtggtgat ggctgtgctg gcggccaaga ctggctacga ggccgacatg gtcgaggcgg acatggacct ggaggccgag ctcggcatcg actcgatcaa gcgcgtggag atcctctcgg aggtgcaggg ccagctgggc gtcgaggcca aggacgtgga tgcgctgagc cgcacgcgca cggtcggtga ggttgtggat gccatgaagg cggaaatcgt ggctgcctct ggtggtagtg ctcctgctgc tgctgttcct tcggcgcccg ctgcttctgc agctccgact cctgcgactg cgccttctgc tgatctgcaa gcgctgctgt ccaaggcgga gactgtggtg atggctgtgc tggcggccaa gactggctac gaggccgaca tggtcgaggc ggacatggac ctggaggccg agctcggcat cgactcgatc aagcgcgtgg agatcctttc cgaggtgcag ggccagctgg gcgtcgaggc caaggacgta gatgcgctga gccgcacgcg cacggtcggt gaagtggtgg acgccatgaa ggcggagatc gtggctgcct ctggtggtag tgctcctgct gctccttcgg cgcccgcgct tcttccaacg ctgtttggtt ccgagtgcga ggacctgtct ctg <210〉 12 <211> 701 <212> PRT <213> 裂殖壺菌物種 <400> 12G 201038734 gtggatgcca tgaaggcgga aatcgtggct gcctctgctg gtagtgctcc tgctcctgct gttccttcgg cgcccgctgc ttctgcagct ccgactcccg ctgcttcgac tgcgccttct gctgatctgc aagcgctgct gtccaaggcg gagacggtgg tgatggctgt gctggcggcc aagactggct acgaggccga catggtcgag gcggacatgg acctggaggc cgagctcggc atcgactcga tcaagcgcgt ggagatcctc tcggaggtgc agggccagct gggcgtcgag gccaaggacg tggatgcgct gagccgcacg cgcacggtcg gtgaggttgt ggatgccatg aaggcggaaa tcgtggctgc ctctggtggt agtgctcctg ctcctgcggt tccttcggcg cccgctgctt ctgcagctcc gactcccgcg gctgcgacag cgccttctgc tgatctgcaa gcgctgctgg ccaaggcgga gactgtggtg atggctgtgc tggcggccaa gactggctac gaggccgaca tggtcgaggc ggacatggac ctggaggccg agctcggcat cgactcgatc aagcgcgtgg agatcctttc cgaggtgcag ggccagctgg gcgtcgaggc caaggacgta gatgcgctga gccgcacgcg cacggtcggt gaggttgtgg atgccatgaa ggcggagatc gtggctgcct ctgctggtag tgctcctgct cctgctgttc cttcggcgcc cgctgcttct gcagctccga ctcccgctgc ttcgactgcg ccttctgctg atctgcaagc gctgctgtcc aaggcggaga ctgtggtgat ggctgtgctg gcggccaaga ctggctacga ggccgacat g gtcgaggcgg acatggacct ggaggccgag ctcggcatcg actcgatcaa gcgcgtggag atcctctcgg aggtgcaggg ccagctgggc gtcgaggcca aggacgtgga tgcgctgagc cgcacgcgca cggtcggtga ggttgtggat gccatgaagg cggaaatcgt ggctgcctct ggtggtagtg ctcctgctgc tgctgttcct tcggcgcccg ctgcttctgc agctccgact cctgcgactg cgccttctgc tgatctgcaa gcgctgctgt ccaaggcgga gactgtggtg atggctgtgc tggcggccaa gactggctac gaggccgaca tggtcgaggc ggacatggac ctggaggccg agctcggcat cgactcgatc aagcgcgtgg agatcctttc cgaggtgcag ggccagctgg gcgtcgaggc caaggacgta gatgcgctga gccgcacgcg cacggtcggt gaagtggtgg Acgccatgaa ggcggagatc gtggctgcct ctggtggtag tgctcctgct gctccttcgg cgcccgcgct tcttccaacg ctgtttggtt ccgagtgcga ggacctgtct ctg <210> 12 <211> 701 <212> PRT <213> Schizochytrium species <400>

Ser Gly Asn Ser Lys Ser Thr Arg Gly Ser Ala Asp Leu Gin Ala Leu 1 5 10 15Ser Gly Asn Ser Lys Ser Thr Arg Gly Ser Ala Asp Leu Gin Ala Leu 1 5 10 15

Leu Ala Lys Ala Glu Thr Val Val Met Ala Val Leu Ala Ala Lys Thr 20 25 30Leu Ala Lys Ala Glu Thr Val Val Met Ala Val Leu Ala Ala Lys Thr 20 25 30

Gly Tyr Glu Ala Asp Met Val Glu Ala Asp Met Asp Leu Glu Ala Glu 35 40 45Gly Tyr Glu Ala Asp Met Val Glu Ala Asp Met Asp Leu Glu Ala Glu 35 40 45

Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin 50 55 60 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2103 35 201038734Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin 50 55 60 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2103 35 201038734

Gly Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr 65 70 75 80Gly Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr 65 70 75 80

Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala Glu He Val Ala 85 90 95Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala Glu He Val Ala 85 90 95

Ala Ser Gly Gly Ser Ala Pro Ala Val Pro Ser Ala Pro Ala Ala Ser 100 105 110Ala Ser Gly Gly Ser Ala Pro Ala Val Pro Ser Ala Pro Ala Ala Ser 100 105 110

Ala Ala Pro Thr Pro Ala Ala Ser Thr Ala Pro Ser Ala Asp Leu Gin 115 120 125Ala Ala Pro Thr Pro Ala Ala Ser Thr Ala Pro Ser Ala Asp Leu Gin 115 120 125

Ala Leu Leu Ser Lys Ala Glu Thr Val Val Met Ala Val Leu Ala Ala 130 135 140Ala Leu Leu Ser Lys Ala Glu Thr Val Val Met Ala Val Leu Ala Ala 130 135 140

Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala Asp Met Asp 145 150 155Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala Asp Met Asp 145 150 155

uo 1 6 G1 u e LUo 1 6 G1 u e L

Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 165 170Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 165 170

I u 1 GI u 1 G

u X G r 5 e 7 s 1 u e Lu X G r 5 e 7 s 1 u e L

Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu Ser 180 185 190Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu Ser 180 185 190

Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala Glu lie 195 200 205Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala Glu lie 195 200 205

a V a V ao 12 A 2 〇 r p a _—_ A 〇 r p a _—_ A r 5 el s 2y _—_ G a i—I A r e s a 1 A a o <—_ I—I A 2 A r e s o r p 〇 r p a •—I A r h T r e s 3 5 13 A 2 a _—I A 〇 r p r h T o r p a o 13 A 2 a «X A r e s a f—I A a _—_ A 0 5 r 2 p 2 r o e 4 s 2a V a V ao 12 A 2 〇rpa ___ A 〇rpa ___ A r 5 el s 2y ___ G ai-IA resa 1 A ao <-_ I-IA 2 A resorp 〇rpa •- IA rh T res 3 5 13 A 2 a _—IA 〇rprh T orpao 13 A 2 a «XA resaf—IA a _—_ A 0 5 r 2 p 2 roe 4 s 2

Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu Thr Val Val 245 250 a _—1 A t 5 e 5 M2Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu Thr Val Val 245 250 a _—1 A t 5 e 5 M2

Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala Asp 260 265 270 Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu 275 280 285 lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp Val 290 295 300Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala Asp 260 265 270 Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu 275 280 285 lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp Val 290 295 300

to e 2 M3To e 2 M3

Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 305 310 315Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 305 310 315

Lys Ala Glu lie Val Ala Ala Ser Gly Gly Ser Ala Pro Ala Pro Ala 325 330 335Lys Ala Glu lie Val Ala Ala Ser Gly Gly Ser Ala Pro Ala Pro Ala 325 330 335

Val Pro Ser Ala Pro Ala Ala Ser Ala Ala Pro Thr Pro Ala Ala Ala 340 345 350Val Pro Ser Ala Pro Ala Ala Ser Ala Ala Pro Thr Pro Ala Ala Ala 340 345 350

Thr Ala Pro Ser Ala Asp Leu Gin Ala Leu Leu Ala Lys Ala Gla Thr 36 201038734 355 360 365Thr Ala Pro Ser Ala Asp Leu Gin Ala Leu Leu Ala Lys Ala Gla Thr 36 201038734 355 360 365

Val Val Met Ala Val Leu Ala Ala 370 375Val Val Met Ala Val Leu Ala Ala 370 375

Lys Thr Gly Tyr Glu Ala Asp 380Lys Thr Gly Tyr Glu Ala Asp 380

Val Glu Ala Asp Met Asp Leu Glu 385 390Val Glu Ala Asp Met Asp Leu Glu 385 390

Ala Glu Leu Gly lie Asp Ser 395Ala Glu Leu Gly lie Asp Ser 395

Lys Arg Val Glu lie Leu Ser Glu 405Lys Arg Val Glu lie Leu Ser Glu 405

Val Gin Gly Gin Leu Gly Val 410 415Val Gin Gly Gin Leu Gly Val 410 415

Ala Lys Asp Val Asp Ala Leu Ser 420Ala Lys Asp Val Asp Ala Leu Ser 420

Arg Thr Arg Thr Val Gly Glu 425 430Arg Thr Arg Thr Val Gly Glu 425 430

Val Asp Ala Met Lys Ala Glu lie 435 440Val Asp Ala Met Lys Ala Glu lie 435 440

Val Ala Ala Ser Ala Gly Ser 445 ΟVal Ala Ala Ser Ala Gly Ser 445 Ο

Pro Ala Pro Ala Val Pro Ser Ala 450 455Pro Ala Pro Ala Val Pro Ser Ala 450 455

Pro Ala Ala Ser Ala Ala Pro 460Pro Ala Ala Ser Ala Ala Pro 460

Pro Ala Ala Ser Thr Ala Pro Ser 465 470Pro Ala Ala Ser Thr Ala Pro Ser 465 470

Ala Asp Leu Gin Ala Leu Leu 475Ala Asp Leu Gin Ala Leu Leu 475

Lys Ala Glu Thr Val Val Met Ala 485Lys Ala Glu Thr Val Val Met Ala 485

Val Leu Ala Ala Lys Thr Gly 490 495Val Leu Ala Ala Lys Thr Gly 490 495

Glu Ala Asp Met Val Glu Ala Asp 500Glu Ala Asp Met Val Glu Ala Asp 500

Met Asp Leu Glu Ala Glu Leu 505 510 lie Asp Ser 工le Lys Arg Val Glu 515 520 lie Leu Ser Glu Val Gin Gly 525Met Asp Leu Glu Ala Glu Leu 505 510 lie Asp Ser Le Lys Arg Val Glu 515 520 lie Leu Ser Glu Val Gin Gly 525

Leu Gly Val Glu Ala Lys Asp Val 530 535Leu Gly Val Glu Ala Lys Asp Val 530 535

Asp Ala Leu Ser Arg Thr Arg 540Asp Ala Leu Ser Arg Thr Arg 540

Val Gly Glu Val Val Asp Ala Met 545 550Val Gly Glu Val Val Asp Ala Met 545 550

Lys Ala Glu lie Val Ala Ala 555Lys Ala Glu lie Val Ala Ala 555

Gly Gly Ser Ala Pro Ala Ala Ala 565Gly Gly Ser Ala Pro Ala Ala Ala 565

Val Pro Ser Ala Pro Ala Ala 570 575Val Pro Ser Ala Pro Ala Ala 570 575

Ala Ala Pro Thr Pro Ala Thr Ala 580Ala Ala Pro Thr Pro Ala Thr Ala 580

Pro Ser Ala Asp Leu Gin Ala 585 590Pro Ser Ala Asp Leu Gin Ala 585 590

Leu Ser Lys Ala Glu Thr Val Val 595 600Leu Ser Lys Ala Glu Thr Val Val 595 600

Met Ala Val Leu Ala Ala Lys 605Met Ala Val Leu Ala Ala Lys 605

Gly Tyr Glu Ala Asp Met Val Glu 610 615Gly Tyr Glu Ala Asp Met Val Glu 610 615

Ala Asp Met Asp Leu Glu Ala 620Ala Asp Met Asp Leu Glu Ala 620

Leu Gly lie Asp Ser lie Lys Arg 625 630Leu Gly lie Asp Ser lie Lys Arg 625 630

Val Glu lie Leu Ser Glu Val 635Val Glu lie Leu Ser Glu Val 635

Gly Gin Leu Gly Val Glu Ala Lys 645Gly Gin Leu Gly Val Glu Ala Lys 645

Asp Val Asp Ala Leu Ser Arg 650 655Asp Val Asp Ala Leu Ser Arg 650 655

Met lie 400 Glu Val Ala Thr Ser 480 Tyr Gly Gin Thr Ser 560 Ser Leu Thr Glu Gin 640 Thr 37 201038734Met lie 400 Glu Val Ala Thr Ser 480 Tyr Gly Gin Thr Ser 560 Ser Leu Thr Glu Gin 640 Thr 37 201038734

Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala Glu lie Val Ala 660 665 670Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala Glu lie Val Ala 660 665 670

Ala Ser Gly Gly Ser Ala Pro Ala Ala Pro Ser Ala Pro Ala Leu Leu 675 680 685Ala Ser Gly Gly Ser Ala Pro Ala Ala Pro Ser Ala Pro Ala Leu Leu 675 680 685

Pro Thr Leu Phe Gly Ser Glu Cys Glu Asp Leu Ser Leu 690 695 700 <210> 13 <211> 276 <212> DNA <213> 裂殖壺菌物種Pro Thr Leu Phe Gly Ser Glu Cys Glu Asp Leu Ser Leu 690 695 700 <210> 13 <211> 276 <212> DNA <213> Schizochytrium species

<210> <211〉 <212> <213〉 種物 菌 壺 ^殖 4 2RJX 1 9 P香 <400> 13 agtgctgatc tgcaagcgct gctggccaag gcggagactg tggtgatggc tgtgctggct 60 gccaagactg gctacgaggc cgacatggtt gaggcggaca tggacctgga ggccgagctc 120 ggcatcgact cgatcaagcg cgtggagatc ctttccgagg tgcagggcca gctgggcgtc 180 gaggccaagg acgtggatgc gctgagccgc acgcgcacgg tcggtgaggt tgtggacgcc 240 atgaaggcgg agatcgtggc tgcctctggt ggtagt 276 <400> 14<210><211><212><213> species bacteria 4 4RJX 1 9 P incense <400> 13 agtgctgatc tgcaagcgct gctggccaag gcggagactg tggtgatggc tgtgctggct 60 gccaagactg gctacgaggc cgacatggtt gaggcggaca tggacctgga ggccgagctc 120 ggcatcgact cgatcaagcg cgtggagatc Ctttccgagg tgcagggcca gctgggcgtc 180 gaggccaagg acgtggatgc gctgagccgc acgcgcacgg tcggtgaggt tgtggacgcc 240 atgaaggcgg agatcgtggc tgcctctggt ggtagt 276 <400> 14

Ser Ala Asp Leu Gin Ala Leu Leu Ala Lys Ala Glu Thr Val Val Met 1 5 10 15Ser Ala Asp Leu Gin Ala Leu Leu Ala Lys Ala Glu Thr Val Val Met 1 5 10 15

Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala 20 25 30Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala 20 25 30

Asp Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 35 40 45Asp Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 35 40 45

Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 50 55 60Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 50 55 60

Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 65 70 75 80Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 65 70 75 80

Met Lys Ala Glu lie Val Ala Ala Ser Gly Gly Ser 85 90 <210〉 15 <211> 276 <212> DNA <213> 裂殖壺菌物種 <400> 15 tctgctgatc tgcaagcgct gctgtccaag gcggagactg tggtgatggc tgtgctggcg 60 gccaagactg gctacgaggc cgacatggtc gaggcggaca tggacctgga ggccgagctc 120 ggcatcgact cgatcaagcg cgtggagatc ctctcggagg tgcagggcca gctgggcgtc 180 gaggccaagg acgtggatgc gctgagccgc acgcgcacgg tcggtgaggt tgtggatgcc 240 atgaaggcgg aaatcgtggc tgcctctgct ggtaqt 276 38 201038734 <210> 16 <211> 92 <212> PRT <213> 裂金壺菌物種 <400> 16Met Lys Ala Glu lie Val Ala Ala Ser Gly Gly Ser 85 90 <210> 15 <211> 276 <212> DNA <213> Schizochytrium species <400> 15 tctgctgatc tgcaagcgct gctgtccaag gcggagactg tggtgatggc tgtgctggcg 60 gccaagactg gctacgaggc cgacatggtc gaggcggaca tggacctgga ggccgagctc 120 ggcatcgact cgatcaagcg cgtggagatc ctctcggagg tgcagggcca gctgggcgtc 180 gaggccaagg acgtggatgc gctgagccgc acgcgcacgg tcggtgaggt tgtggatgcc 240 atgaaggcgg aaatcgtggc tgcctctgct ggtaqt 276 38 201038734 < 210 > 16 < 211 > 92 < 212 > PRT < 213 > split pot of gold Fungus species <400> 16

Ser Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu Thr Val Val Met 15 10 15Ser Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu Thr Val Val Met 15 10 15

Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala 20 25 30Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala 20 25 30

Asp Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 35 40 45Asp Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 35 40 45

Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 50 55 60Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 50 55 60

Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 65 70 75 80Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 65 70 75 80

Met Lys Ala Glu lie Val Ala Ala Ser Ala Gly Ser 85 90 <210〉 17 <211> 276 <212> DNA <213〉 裂殖壺菌物種 <400〉 17 60 120 180 240 27 6 tctgctgatc tgcaagcgct gctgtccaag gcggagacgg tggtgatggc tgtgctggcg gccaagactg gctacgaggc cgacatggtc gaggcggaca tggacctgga ggccgagctc ggcatcgact cgatcaagcg cgtggagatc ctctcggagg tgcagggcca gctgggcgtc gaggccaagg acgtggatgc gctgagccgc acgcgcacgg tcggtgaggt tgtggatgcc atgaaggcgg aaatcgtggc tgcctctggt ggtagt <210> 18 <211> 92 <212> PRT <213> 裂殖壺菌物種 <400> 18Met Lys Ala Glu lie Val Ala Ala Ser Ala Gly Ser 85 90 <210> 17 <211> 276 <212> DNA <213> Schizochytrium species <400> 17 60 120 180 240 27 6 tctgctgatc tgcaagcgct gctgtccaag gcggagacgg tggtgatggc tgtgctggcg gccaagactg gctacgaggc cgacatggtc gaggcggaca tggacctgga ggccgagctc ggcatcgact cgatcaagcg cgtggagatc ctctcggagg tgcagggcca gctgggcgtc gaggccaagg acgtggatgc gctgagccgc acgcgcacgg tcggtgaggt tgtggatgcc atgaaggcgg aaatcgtggc tgcctctggt ggtagt < 210 > 18 < 211 > 92 < 212 > PRT < 213 > Schizochytrium Species <400> 18

Ser Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu Thr Val Val Met 15 10 15Ser Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu Thr Val Val Met 15 10 15

Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala 20 25 30Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala 20 25 30

Asp Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 35 40 45Asp Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 35 40 45

Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 50 55 60Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 50 55 60

Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 65 70 75 80 39 201038734Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 65 70 75 80 39 201038734

Met Lys Ala Glu 工le Val Ala Ala Ser Gly Gly Ser 85 90 <210> 19 <211> 276 <212〉 DNA <213> 裂殖壺菌物種 <400> 19 tctgctgatc tgcaagcgct gctggccaag gcggagactg tggtgatggc tgtgctggcg 60 gccaagactg gctacgaggc cgacatggtc gaggcggaca tggacctgga ggccgagctc 120 ggcatcgact cgatcaagcg cgtggagatc ctttccgagg tgcagggcca gctgggcgtc 180 gaggccaagg acgtagatgc gctgagccgc acgcgcacgg tcggtgaggt tgtggatgcc 240 atgaaggcgg agatcgtggc tgcctctgct ggtagt 276Met Lys Ala Glu work le Val Ala Ala Ser Gly Gly Ser 85 90 <210> 19 <211> 276 <212> DNA <213> Schizochytrium species <400> 19 tctgctgatc tgcaagcgct gctggccaag gcggagactg tggtgatggc tgtgctggcg 60 gccaagactg gctacgaggc cgacatggtc gaggcggaca tggacctgga ggccgagctc 120 ggcatcgact cgatcaagcg cgtggagatc ctttccgagg tgcagggcca gctgggcgtc 180 gaggccaagg acgtagatgc gctgagccgc acgcgcacgg tcggtgaggt tgtggatgcc 240 atgaaggcgg agatcgtggc tgcctctgct ggtagt 276

<210〉 20 <211> 92 <212> PRT <213> 裂殖壺菌物種 <400> 20<210> 20 <211> 92 <212> PRT <213> Schizochytrium species <400> 20

Ser Ala Asp Leu Gin Ala Leu Leu Ala Lys Ala Glu Thr Val Val Met 15 10 15Ser Ala Asp Leu Gin Ala Leu Leu Ala Lys Ala Glu Thr Val Val Met 15 10 15

Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala 20 25 30Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala 20 25 30

Asp Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 35 40 45Asp Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 35 40 45

Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 50 55 60Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 50 55 60

Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 65 70 75 80Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 65 70 75 80

Met Lys Ala Glu lie Val Ala Ala Ser Ala Gly Ser 85 90 <210> 21 <211> 276 <212> DNA <213〉 裂i壺菌物種 <400> 21 tctgctgatc tgcaagcgct gctgtccaag gcggagactg tggtgatggc tgtgctggcg 60 gccaagactg gctacgaggc cgacatggtc gaggcggaca tggacctgga ggccgagctc 120 ggcatcgact cgatcaagcg cgtggagatc ctctcggagg tgcagggcca gctgggcgtc 180 gaggccaagg acgtggatgc gctgagccgc acgcgcacgg tcggtgaggt tgtggatgcc 240 atgaaggcgg aaatcgtggc tgcctctggt ggtagt 27 6 <210> 22 <211> 92 <212> PRT <213> 裂殖壺菌物種 40 201038734 <400> 22Met Lys Ala Glu lie Val Ala Ala Ser Ala Gly Ser 85 90 <210> 21 <211> 276 <212> DNA <213> Schizochytrium species <400> 21 tctgctgatc tgcaagcgct gctgtccaag gcggagactg tggtgatggc tgtgctggcg 60 gccaagactg gctacgaggc cgacatggtc gaggcggaca tggacctgga ggccgagctc 120 ggcatcgact cgatcaagcg cgtggagatc ctctcggagg tgcagggcca gctgggcgtc 180 gaggccaagg acgtggatgc gctgagccgc acgcgcacgg tcggtgaggt tgtggatgcc 240 atgaaggcgg aaatcgtggc tgcctctggt ggtagt 27 6 < 210 > 22 < 211 > 92 < 212 > PRT < 213 > Schizochytrium Species 40 201038734 <400> 22

Ser Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu Thr Val Val Met 15 10 15Ser Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu Thr Val Val Met 15 10 15

Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala 20 25 30Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala 20 25 30

Asp Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 35 40 45Asp Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 35 40 45

Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 50 55 60Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 50 55 60

Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 65 70 75 80Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 65 70 75 80

Met Lys Ala Glu lie Val Ala Ala Ser Gly Gly Ser 85 90 <210> 23 <211> 276 <212> DNA <213> 裂殖壺菌物種 <40〇> 23 60 120 180 240 276 tctgctgatc tgcaagcgct gctgtccaag gcggagactg tggtgatggc tgtgctggcg gccaagactg gctacgaggc cgacatggtc gaggcggaca tggacctgga ggccgagctc ggcatcgact cgatcaagcg cgtggagatc ctttccgagg tgcagggcca gctgggcgtc gaggccaagg acgtagatgc gctgagccgc acgcgcacgg tcggtgaagt ggtggacgcc atgaaggcgg agatcgtggc tgcctctggt ggtagt <210> 24 <211> 92 <212> PRT <213> 裂殖壺菌物種 <400> 24Met Lys Ala Glu lie Val Ala Ala Ser Gly Gly Ser 85 90 <210> 23 <211> 276 <212> DNA <213> Schizochytrium species <40〇> 23 60 120 180 240 276 tctgctgatc tgcaagcgct gctgtccaag gcggagactg tggtgatggc tgtgctggcg gccaagactg gctacgaggc cgacatggtc gaggcggaca tggacctgga ggccgagctc ggcatcgact cgatcaagcg cgtggagatc ctttccgagg tgcagggcca gctgggcgtc gaggccaagg acgtagatgc gctgagccgc acgcgcacgg tcggtgaagt ggtggacgcc atgaaggcgg agatcgtggc tgcctctggt ggtagt < 210 > 24 < 211 > 92 < 212 > PRT < 213 > Schizochytrium Fungus species <400> 24

Ser Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu Thr Val Val Met 15 10 15Ser Ala Asp Leu Gin Ala Leu Leu Ser Lys Ala Glu Thr Val Val Met 15 10 15

Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala 20 25 30Ala Val Leu Ala Ala Lys Thr Gly Tyr Glu Ala Asp Met Val Glu Ala 20 25 30

Asp Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 35 40 45Asp Met Asp Leu Glu Ala Glu Leu Gly lie Asp Ser lie Lys Arg Val 35 40 45

Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 50 55 60Glu lie Leu Ser Glu Val Gin Gly Gin Leu Gly Val Glu Ala Lys Asp 50 55 60

Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 65 7 0 75 80Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala 65 7 0 75 80

Met Lys Ala Glu lie Val Ala Ala Ser Gly Gly Ser 85 90 41 201038734 <210> 25 <211> 2178 <212> DNA <213> 裂殖壺菌物種 <400〉 25 gcgctacagg cggcgctcac gtccgtcgag gcgcagttcg gcaaggtggg tggctttgtg 60 ttccagttcg gcgacgacga cgtgcaagcg cagctcggct gggcgctgct cgcggccaag 120 cacctcaaaa cttcgctgtc agaacagatc gagggcggtc gcaccttttt cgtggccgtc 180 gcgcggctcg acggccagct ggggctctcc ggcaagtcga cgaccgctac cgttgatctc 240 tcccgcgcgc sgc^gggcag cgtgttcggc ctgtgcaaga cactcgacct ggagtggccc 300 gctgtcttct gccgcggaat cgacctggcc gccgacctcg acgccgcaca ggccgcgcgg 360 tgcctgctgg gcgagctgtc agaccccgac gtggccgtgc gcgagtctgg ttactccgcc 420 tcgggccagc gctgcacgac aactacgaag tcgctgacta cgggcaagcc gcaccagccg 480 atctcctcgt cggacctctt tctggtgtcg ggcggcgcgc gcggcatcac cccgctgtgc 540 gtgcgcgagc tggcgcagcg cgtgggcggc ggcacgtacg tgctcatcgg ccgctcggag 600 ctgcccacga cggagcctgc ctgggcggtc ggcgtggagt ctggcaagcc gctggagaag 660 gccgcgctgg cgttcctgaa ggcggagttt gcagcgggcc gcggggccaa gccgacgccg 720 atgctgcaca agaagctcgt gggcgccgtg gtcggagcgc gcgaggtgcg agcctcgctc 780 gccgagatca ctgcacaggg cgccacggct gtgtacgagt cgtgcgacgt gagctctgcc 840 gccaaggtgc gtgagatggt agagcgcgtg cagcagcagg gcgggcggcg cgtgtcgggc 900 gtgttccacg cgtcgggcgt gctgcgcgac aagctcgtgg agaacaagtc gctggcggac 960 ttcagcgccg tgtacgacac caaggtgggc ggcctcatca acctgctggc ctgcgtggac 1020 ctggcgcagc tgcgtcacct cgtgctcttc agctcgctcg cgggcttcca cggcaacgtc 1080 gggcagtcgg actacgcaat ggccaacgag gcgctcaaca agctggcggc gcacctgtcg 1140 gcggtgcacc cgcagctgtg cgcgcgctcg atctgcttcg gaccgtggga cggcggcatg 1200 gtgacccccg cgctcaaggc caacttcatc cgcatgggca tccagatcat cccgcgccaa 1260 ggcggcgcgc agaccgtcgc caacatgctc gtcagtagct cccccggtca gctgctcgtg 1320 ggcaactggg gcgtgccacc cgtcgtgccg agtgccaccg agcacaccgt gctgcagacg 1380 ctccgccaga gcgacaaccc cttcctcgac tcgcacgtga tccagggccg ccgcgtgctg 1440 cccatgaccc tggccgtggg ctacatggcg caccaggcgc agagcatcta cgcgggccac 1500 cagctgtggg ccgtcgagga cgcccagctc ttcaagggca tcgccatcga caatggcgcc 1560 gacgtgcccg tgcgcgtgga gctgtcgcgc cgcaaggagg agcaggagga cgccggcaag 1620 gtcaaggtca aggtgcaggt gctgctcaaa tcgcaggtca acggcaagtc ggtgcccgcg 1680 tacaaggcga ccgtcgtgct gtcccctgcg ccgcgcccca gcgtcatcac gcgtgacttc 1740 gacctcaccc cggacccggc ctgcacggag cacgacctct acgacggcaa gacgctcttc 1800 cacggcaagg ccttccaggg catcgagcag gtgctctcgg cgacgcccaa gcagctcacc 1860 gccaagtgcc gcaatttgcc cctcacgccc gagcagcgcg gccagttcgt cgttaacctc 1920 agccagcagg acccgttcca ggcggacatt gcgttccagg cgatgctcgt ctgggcgcgc 1980 atgctgcgcc aatcggcggc cctgcccaac aactgcgagc gcttcgactt ttacaagccg 2040 atggccccgg gcgccaccta ctacacgtcg gtcaagctgg cctcggcctc acccttggtg 2100Met Lys Ala Glu lie Val Ala Ala Ser Gly Gly Ser 85 90 41 201038734 <210> 25 <211> 2178 <212> DNA <213> Schizochytrium species <400> 25 gcgctacagg cggcgctcac gtccgtcgag gcgcagttcg gcaaggtggg tggctttgtg 60 ttccagttcg gcgacgacga cgtgcaagcg cagctcggct gggcgctgct cgcggccaag 120 cacctcaaaa cttcgctgtc agaacagatc gagggcggtc gcaccttttt cgtggccgtc 180 gcgcggctcg acggccagct ggggctctcc ggcaagtcga cgaccgctac cgttgatctc 240 tcccgcgcgc sgc ^ gggcag cgtgttcggc ctgtgcaaga cactcgacct ggagtggccc 300 gctgtcttct gccgcggaat cgacctggcc gccgacctcg acgccgcaca ggccgcgcgg 360 tgcctgctgg gcgagctgtc agaccccgac gtggccgtgc gcgagtctgg ttactccgcc 420 tcgggccagc gctgcacgac aactacgaag tcgctgacta cgggcaagcc gcaccagccg 480 atctcctcgt cggacctctt tctggtgtcg ggcggcgcgc gcggcatcac cccgctgtgc 540 gtgcgcgagc tggcgcagcg cgtgggcggc ggcacgtacg tgctcatcgg ccgctcggag 600 ctgcccacga cggagcctgc ctgggcggtc ggcgtggagt ctggcaagcc gctggagaag 660 gccgcgctgg cgttcctgaa ggcggagttt gcagcgggcc gcggggccaa gcc gacgccg 720 atgctgcaca agaagctcgt gggcgccgtg gtcggagcgc gcgaggtgcg agcctcgctc 780 gccgagatca ctgcacaggg cgccacggct gtgtacgagt cgtgcgacgt gagctctgcc 840 gccaaggtgc gtgagatggt agagcgcgtg cagcagcagg gcgggcggcg cgtgtcgggc 900 gtgttccacg cgtcgggcgt gctgcgcgac aagctcgtgg agaacaagtc gctggcggac 960 ttcagcgccg tgtacgacac caaggtgggc ggcctcatca acctgctggc ctgcgtggac 1020 ctggcgcagc tgcgtcacct cgtgctcttc agctcgctcg cgggcttcca cggcaacgtc 1080 gggcagtcgg actacgcaat ggccaacgag gcgctcaaca agctggcggc gcacctgtcg 1140 gcggtgcacc cgcagctgtg cgcgcgctcg atctgcttcg gaccgtggga cggcggcatg 1200 gtgacccccg cgctcaaggc caacttcatc cgcatgggca tccagatcat cccgcgccaa 1260 ggcggcgcgc agaccgtcgc caacatgctc gtcagtagct cccccggtca gctgctcgtg 1320 ggcaactggg gcgtgccacc cgtcgtgccg agtgccaccg agcacaccgt gctgcagacg 1380 ctccgccaga gcgacaaccc cttcctcgac tcgcacgtga tccagggccg ccgcgtgctg 1440 cccatgaccc tggccgtggg ctacatggcg caccaggcgc agagcatcta cgcgggccac 1500 cagctgtggg ccgtcgagga cgcccagctc ttcaagggca tcgccatcga caatggcgcc 1560 gacgtgcccg tgcgcgtgga gctgtcgcgc cgcaaggagg agcaggagga cgccggcaag 1620 gtcaaggtca aggtgcaggt gctgctcaaa tcgcaggtca acggcaagtc ggtgcccgcg 1680 tacaaggcga ccgtcgtgct gtcccctgcg ccgcgcccca gcgtcatcac gcgtgacttc 1740 gacctcaccc cggacccggc ctgcacggag cacgacctct acgacggcaa gacgctcttc 1800 cacggcaagg ccttccaggg catcgagcag gtgctctcgg cgacgcccaa gcagctcacc 1860 gccaagtgcc gcaatttgcc cctcacgccc gagcagcgcg gccagttcgt cgttaacctc 1920 agccagcagg acccgttcca ggcggacatt gcgttccagg cgatgctcgt ctgggcgcgc 1980 atgctgcgcc aatcggcggc cctgcccaac aactgcgagc gcttcgactt ttacaagccg 2040 atggccccgg gcgccaccta ctacacgtcg gtcaagctgg cctcggcctc acccttggtg 2100

42 201038734 gactctgtgt gcaagtgcac cgtggcgatg cacgatgagc aaggtgaggt gtacttttct 2160 gctcgtgcca gcgtcgtc 2178 <210> 26 <211> 726 <212> PRT <213> 裂殖壺菌物種 <400> 2642 201038734 gactctgtgt gcaagtgcac cgtggcgatg cacgatgagc aaggtgaggt gtacttttct 2160 gctcgtgcca gcgtcgtc 2178 <210> 26 <211> 726 <212> PRT <213> Schizochytrium species <400>

Ala Leu Gin Ala Ala Leu Thr Ser Val Glu Ala Gin Phe Gly Lys Val 15 10 15Ala Leu Gin Ala Ala Leu Thr Ser Val Glu Ala Gin Phe Gly Lys Val 15 10 15

Gly Gly Phe Val Phe Gin Phe Gly Asp Asp Asp Val Gin Ala Gin Leu 20 25 30Gly Gly Phe Val Phe Gin Phe Gly Asp Asp Asp Val Gin Ala Gin Leu 20 25 30

Gly Trp Ala Leu Leu Ala Ala Lys His Leu Lys Thr Ser Leu Ser Glu 35 40 45 o 〇Gly Trp Ala Leu Leu Ala Ala Lys His Leu Lys Thr Ser Leu Ser Glu 35 40 45 o 〇

Gin lie Glu Gly Gly Arg Thr Phe Phe Val Ala Val Ala Arg Leu Asp 50 55 60Gin lie Glu Gly Gly Arg Thr Phe Phe Val Ala Val Ala Arg Leu Asp 50 55 60

Gly Gin Leu Gly Leu Ser Gly Lys Ser Thr Thr Ala Thr Val Asp Leu 65 70 75 80Gly Gin Leu Gly Leu Ser Gly Lys Ser Thr Thr Ala Thr Val Asp Leu 65 70 75 80

Ser Arg Ala Gin Gin Gly Ser Val Phe Gly Leu Cys Lys Thr Leu Asp 85 90 95Ser Arg Ala Gin Gin Gly Ser Val Phe Gly Leu Cys Lys Thr Leu Asp 85 90 95

Leu Glu Trp Pro Ala Val Phe Cys Arg Gly lie Asp Leu Ala Ala Asp 100 105 110Leu Glu Trp Pro Ala Val Phe Cys Arg Gly lie Asp Leu Ala Ala Asp 100 105 110

Leu Asp Ala Ala Gin Ala Ala Arg Cys Leu Leu Gly Glu Leu Ser Asp 115 120 125Leu Asp Ala Ala Gin Ala Ala Arg Cys Leu Leu Gly Glu Leu Ser Asp 115 120 125

Pro Asp Val Ala Val Arg Glu Ser Gly Tyr Ser Ala Ser Gly Gin Arg 130 135 140Pro Asp Val Ala Val Arg Glu Ser Gly Tyr Ser Ala Ser Gly Gin Arg 130 135 140

Cys Thr Thr Thr Thr Lys Ser Leu Thr Thr Gly Lys Pro His Gin Pro 145 150 155 160 lie Ser Ser Ser Asp Leu Phe Leu Val Ser Gly Gly Ala Arg Gly lie 165 170 175Cys Thr Thr Thr Thr Lys Ser Leu Thr Thr Gly Lys Pro His Gin Pro 145 150 155 160 lie Ser Ser Ser Asp Leu Phe Leu Val Ser Gly Gly Ala Arg Gly lie 165 170 175

Thr Pro Leu Cys Val Arg Glu Leu Ala Gin Arg Val Gly Gly Gly Thr 180 185 190Thr Pro Leu Cys Val Arg Glu Leu Ala Gin Arg Val Gly Gly Gly Thr 180 185 190

Tyr Val Leu lie Gly Arg Ser Glu Leu Pro Thr Thr Glu Pro Ala Trp 195 200 205Tyr Val Leu lie Gly Arg Ser Glu Leu Pro Thr Thr Glu Pro Ala Trp 195 200 205

Ala Val Gly Val Glu Ser Gly Lys Pro Leu Glu Lys Ala Ala Leu Ala 210 215 220Ala Val Gly Val Glu Ser Gly Lys Pro Leu Glu Lys Ala Ala Leu Ala 210 215 220

Phe Leu Lys Ala Glu Phe Ala Ala Gly Arg Gly Ala Lys Pro Thr Pro 225 230 235 240Phe Leu Lys Ala Glu Phe Ala Ala Gly Arg Gly Ala Lys Pro Thr Pro 225 230 235 240

s 5 y4 L 2 s Ly s •1 H u e L t e Ms 5 y4 L 2 s Ly s •1 H u e L t e M

Leu Val Gly Ala Val Val Gly Ala Arg Glu Val 250 255 43 201038734Leu Val Gly Ala Val Val Gly Ala Arg Glu Val 250 255 43 201038734

Arg Ala Ser Leu Ala Glu lie Thr Ala Gin Gly Ala Thr Ala Val Tyr 260 265 270Arg Ala Ser Leu Ala Glu lie Thr Ala Gin Gly Ala Thr Ala Val Tyr 260 265 270

Glu Ser Cys Asp Val Ser Ser Ala Ala Lys Val Arg Glu Met Val Glu 275 280 285Glu Ser Cys Asp Val Ser Ser Ala Ala Lys Val Arg Glu Met Val Glu 275 280 285

Arg Val Gin Gin Gin Gly Gly Arg Arg Val Ser Gly Val Phe His Ala 290 295 300Arg Val Gin Gin Gin Gly Gly Arg Arg Val Ser Gly Val Phe His Ala 290 295 300

Ser Gly Val Leu Arg Asp Lys Leu Val Glu Asn Lys Ser Leu Ala Asp 305 310 315 320Ser Gly Val Leu Arg Asp Lys Leu Val Glu Asn Lys Ser Leu Ala Asp 305 310 315 320

Phe Ser Ala Val Tyr Asp Thr Lys Val Gly Gly Leu lie Asn Leu Leu 325 330 335Phe Ser Ala Val Tyr Asp Thr Lys Val Gly Gly Leu lie Asn Leu Leu 325 330 335

Ala Cys Val Asp Leu Ala Gin Leu Arg His Leu Val Leu Phe Ser Ser 340 345 350Ala Cys Val Asp Leu Ala Gin Leu Arg His Leu Val Leu Phe Ser Ser 340 345 350

Leu Ala Gly Phe His Gly Asn Val Gly Gin Ser Asp Tyr Ala Met Ala 355 360 365Leu Ala Gly Phe His Gly Asn Val Gly Gin Ser Asp Tyr Ala Met Ala 355 360 365

Asn Glu Ala Leu Asn Lys Leu Ala Ala His Leu Ser Ala Val His Pro 370 375 380Asn Glu Ala Leu Asn Lys Leu Ala Ala His Leu Ser Ala Val His Pro 370 375 380

Gin Leu Cys Ala Arg Ser lie Cys Phe Gly Pro Trp Asp Gly Gly Met 385 390 395 400Gin Leu Cys Ala Arg Ser lie Cys Phe Gly Pro Trp Asp Gly Gly Met 385 390 395 400

Val Thr Pro Ala Leu Lys Ala Asn Phe lie Arg Met Gly lie Gin lie 405 410 415 lie Pro Arg Gin Gly Gly Ala Gin Thr Val Ala Asn Met Leu Val Ser 420 425 430Val Thr Pro Ala Leu Lys Ala Asn Phe lie Arg Met Gly lie Gin lie 405 410 415 lie Pro Arg Gin Gly Gly Ala Gin Thr Val Ala Asn Met Leu Val Ser 420 425 430

Ser Ser Pro Gly Gin Leu Leu Val Gly Asn Trp Gly Val Pro Pro Val 435 440 445Ser Ser Pro Gly Gin Leu Leu Val Gly Asn Trp Gly Val Pro Pro Val 435 440 445

Val Pro Ser Ala Thr Glu His Thr Val Leu Gin Thr Leu Arg Gin Ser 450 455 460Val Pro Ser Ala Thr Glu His Thr Val Leu Gin Thr Leu Arg Gin Ser 450 455 460

Asp Asn Pro Phe Leu Asp Ser His Val lie Gin Gly Arg Arg Val Leu 465 470 475 480Asp Asn Pro Phe Leu Asp Ser His Val lie Gin Gly Arg Arg Val Leu 465 470 475 480

Pro Met Thr Leu Ala Val Gly Tyr Met Ala His Gin Ala Gin Ser lie 485 490 495Pro Met Thr Leu Ala Val Gly Tyr Met Ala His Gin Ala Gin Ser lie 485 490 495

Tyr Ala Gly His Gin Leu Trp Ala Val Glu Asp Ala Gin Leu Phe Lys 500 505 510Tyr Ala Gly His Gin Leu Trp Ala Val Glu Asp Ala Gin Leu Phe Lys 500 505 510

Gly lie Ala lie Asp Asn Gly Ala Asp Val Pro Val Arg Val Glu Leu 515 520 525Gly lie Ala lie Asp Asn Gly Ala Asp Val Pro Val Arg Val Glu Leu 515 520 525

Ser Arg Arg Lys Glu Glu Gin Glu Asp Ala Gly Lys Val Lys Val Lys 530 535 540Ser Arg Arg Lys Glu Glu Gin Glu Asp Ala Gly Lys Val Lys Val Lys 530 535 540

Val Gin Val Leu Leu Lys Ser Gin Val Asn Gly Lys Ser Val Pro Ala 44 201038734 545 550 555 560Val Gin Val Leu Leu Lys Ser Gin Val Asn Gly Lys Ser Val Pro Ala 44 201038734 545 550 555 560

Tyr Lys Ala Thr Val Val Leu Ser Pro Ala Pro Arg Pro Ser Val lie 565 570 575Tyr Lys Ala Thr Val Val Leu Ser Pro Ala Pro Arg Pro Ser Val lie 565 570 575

Thr Arg Asp Phe Asp Leu Thr Pro Asp Pro Ala Cys Thr Glu His Asp 580 585 590Thr Arg Asp Phe Asp Leu Thr Pro Asp Pro Ala Cys Thr Glu His Asp 580 585 590

Leu Tyr Asp Gly Lys Thr Leu Phe His Gly Lys Ala Phe Gin Gly lie 595 600 605Leu Tyr Asp Gly Lys Thr Leu Phe His Gly Lys Ala Phe Gin Gly lie 595 600 605

Glu Gin Val Leu Ser Ala Thr Pro Lys Gin Leu Thr Ala Lys Cys Arg 610 615 620Glu Gin Val Leu Ser Ala Thr Pro Lys Gin Leu Thr Ala Lys Cys Arg 610 615 620

Asn Leu Pro Leu Thr Pro Glu Gin Arg Gly Gin Phe Val Val Asn Leu 625 630 635 640Asn Leu Pro Leu Thr Pro Glu Gin Arg Gly Gin Phe Val Val Asn Leu 625 630 635 640

Ser Gin Gin Asp Pro Phe Gin Ala Asp lie Ala Phe Gin Ala Met Leu 645 650 655Ser Gin Gin Asp Pro Phe Gin Ala Asp lie Ala Phe Gin Ala Met Leu 645 650 655

Val Trp Ala Arg Met Leu Arg Gin Ser Ala Ala Leu Pro Asn Asn Cys 660 665 670Val Trp Ala Arg Met Leu Arg Gin Ser Ala Ala Leu Pro Asn Asn Cys 660 665 670

Glu Arg Phe Asp Phe Tyr Lys Pro Met Ala Pro Gly Ala Thr Tyr Tyr 675 680 685Glu Arg Phe Asp Phe Tyr Lys Pro Met Ala Pro Gly Ala Thr Tyr Tyr 675 680 685

Thr Ser Val Lys Leu Ala Ser Ala Ser Pro Leu Val Asp Ser Val Cys 690 695 700Thr Ser Val Lys Leu Ala Ser Ala Ser Pro Leu Val Asp Ser Val Cys 690 695 700

Lys Cys Thr Val Ala Met His Asp Glu Gin Gly Glu Val Tyr Phe Ser 705 710 715 720Lys Cys Thr Val Ala Met His Asp Glu Gin Gly Glu Val Tyr Phe Ser 705 710 715 720

Ala Arg Ala Ser Val Val 725 <210> 27 <211〉 39 <212> DNA <213> 裂殖壺菌物種 <400> 27 39 ctcgactcgc acgtgatcca gggccgccgc gtgctgccc <210> 28 <211> 13 <212> PRT <213> 裂殖壺菌物種 <400> 28Ala Arg Ala Ser Val Val 725 <210> 27 <211> 39 <212> DNA <213> Schizochytrium species <400> 27 39 ctcgactcgc acgtgatcca gggccgccgc gtgctgccc <210> 28 <211> 13 <212> PRT <213> Schizochytrium species <400> 28

Leu Asp Ser His Val lie Gin Gly Arg Arg Val Leu Pro 1 5 10 <210> 29 <211> 1341 <212> DNA <213> 裂殖壺菌物種 <400> 29 gataacattg cggtcgtggg catggcggtg cagtatgccg gatgcaagaa ccaggacgag 60 45 201038734 ttctgggata ggtacgcgct tgcaacgatc gacctggccc gccaacctac cagggcgagc ttccgcgact ccgcgcgtgt cgctacagcc cacttgctct ttcttcattc aacccactgt gccatcatgg accttgctcg ccgagcgaga gtgcagtacg gcgctgcgcc ggcaactttg tcgatgcagc ccgctcgtcg aaaccaggcg gcgcactgtg cgctgatgcg accgcgacct gctacggctg ggcgcgccct gcgacttcgg tgctcaatct cgcgcccctg actccgaccc tcgatgcagc cgcgcagcgc tctcggggtt ccgtgccgct tgctgaagcg gcacgagtct agtcgtgcat tggagtgcca actgctttcg gccacactct acggcacgat tggacgaggc atgagctcaa tcttccgtga taaggagatc ccacttccac cgtcgatgcc gctcgacgcc catcgtgagc gtaccaagtg gtcggagcgc ggcgtccttc ctgcgcgtcg ggacgtgatg ctccaccttc gcggcagggc cctcgaggac gagcaacgcc ggaggacctg cgccacgggc aggtaacacg cgtggcggcc cccgcccacg catcccttgg gtgcgcctcg g aactcgagcc ccgcagcgca agcgtcgaca ggaattaacc ggctgcctgt catgtggaga ccgcgcgctg gtggccaacc gcgctgtact ctgtgcggcg caggcgatgc agccagggcc gccgtgcgcg gggtgcggcc tacacgagcg actccgcagg gaccacccgc gggttcgcag cccggtgtcg ccgtactcgt ctctccgcct cgatctcggc gcaagtacgc acgagcacga tcgacgacgc cgttccccat accgcgtggg tctcgcccga agctcggcct gcctcaagct ccacatgctt cgctgggcgg tgacgcccgg acggcgaccg tgccgctgag tcggcatcga gcgacgtcgt cgcgcatggg gcatggccaa accgctccaa cggcgcaggc ttggctttgg ggagcgcctc cgacaccttc cctcctcgcc cagcaccacc ggacaatctg cgcccagcgc ggccagcgac ggggcccgtg ggcgtccgac tccggacccg accggacgat agagggcggc catctacggc cccgcacctg cccaagcgag ggaggtagag ctccaccaag ggtgctgctg ctgcatcgac gcgggcaggc tggaaccaac 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1341Leu Asp Ser His Val lie Gin Gly Arg Arg Val Leu Pro 1 5 10 <210> 29 <211> 1341 <212> DNA <213> Schizochytrium species <400> 29 gataacattg cggtcgtggg catggcggtg cagtatgccg gatgcaagaa ccaggacgag 60 45 201038734 ttctgggata ggtacgcgct tgcaacgatc gacctggccc gccaacctac cagggcgagc ttccgcgact ccgcgcgtgt cgctacagcc cacttgctct ttcttcattc aacccactgt gccatcatgg accttgctcg ccgagcgaga gtgcagtacg gcgctgcgcc ggcaactttg tcgatgcagc ccgctcgtcg aaaccaggcg gcgcactgtg cgctgatgcg accgcgacct gctacggctg ggcgcgccct gcgacttcgg tgctcaatct cgcgcccctg actccgaccc tcgatgcagc cgcgcagcgc tctcggggtt ccgtgccgct tgctgaagcg gcacgagtct agtcgtgcat tggagtgcca actgctttcg gccacactct acggcacgat tggacgaggc atgagctcaa tcttccgtga taaggagatc ccacttccac cgtcgatgcc gctcgacgcc catcgtgagc gtaccaagtg gtcggagcgc ggcgtccttc ctgcgcgtcg ggacgtgatg ctccaccttc gcggcagggc cctcgaggac gagcaacgcc ggaggacctg cgccacgggc aggtaacacg cgtggcggcc cccgcccacg catcccttgg gtgcgcctcg g aactcgagcc ccgcagcgca agcgtcgaca ggaa ttaacc ggctgcctgt catgtggaga ccgcgcgctg gtggccaacc gcgctgtact ctgtgcggcg caggcgatgc agccagggcc gccgtgcgcg gggtgcggcc tacacgagcg actccgcagg gaccacccgc gggttcgcag cccggtgtcg ccgtactcgt ctctccgcct cgatctcggc gcaagtacgc acgagcacga tcgacgacgc cgttccccat accgcgtggg tctcgcccga agctcggcct gcctcaagct ccacatgctt cgctgggcgg tgacgcccgg acggcgaccg tgccgctgag tcggcatcga gcgacgtcgt cgcgcatggg gcatggccaa accgctccaa cggcgcaggc ttggctttgg ggagcgcctc cgacaccttc cctcctcgcc cagcaccacc ggacaatctg cgcccagcgc ggccagcgac ggggcccgtg ggcgtccgac tccggacccg accggacgat Agagggcggc catctacggc cccgcacctg cccaagcgag ggaggtagag ctccaccaag ggtgctgctg ctgcatcgac gcgggcaggc tggaaccaac 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1341

<21〇> 30 <211> 447 <212> PRT <213> 裂殖壺菌物種<21〇> 30 <211> 447 <212> PRT <213> Schizochytrium species

<400> 30<400> 30

Asp Asn lie Ala Val Val Gly Met Ala Val Gin Tyr Ala Gly Cys Lys 15 10 15Asp Asn lie Ala Val Val Gly Met Ala Val Gin Tyr Ala Gly Cys Lys 15 10 15

Asn Gin Asp Glu Phe Trp Asp Thr Leu Met Arg Lys Glu lie Asn Ser 20 25 30Asn Gin Asp Glu Phe Trp Asp Thr Leu Met Arg Lys Glu lie Asn Ser 20 25 30

Ser Pro lie Ser Ala Glu Arg Leu Gly Thr Arg Tyr Arg Asp Leu 35 40 45Ser Pro lie Ser Ala Glu Arg Leu Gly Thr Arg Tyr Arg Asp Leu 35 40 45

Phe His Pro Gin Arg Ser Lys Tyr Ala Asp Thr Phe Cys Asn Asp Arg 50 55 60Phe His Pro Gin Arg Ser Lys Tyr Ala Asp Thr Phe Cys Asn Asp Arg 50 55 60

Tyr Gly Cys Val Asp Ala Ser Val Asp Asn Glu His Asp Leu Leu Ala 65 70 75 80Tyr Gly Cys Val Asp Ala Ser Val Asp Asn Glu His Asp Leu Leu Ala 65 70 75 80

Asp Leu Ala Arg Arg Ala Leu Leu Asp Ala Gly lie Asn Leu Asp Asp 85 90 95 46 201038734Asp Leu Ala Arg Arg Ala Leu Leu Asp Ala Gly lie Asn Leu Asp Asp 85 90 95 46 201038734

Ala Ser Thr Thr Ala Asn Leu Arg 100 Asp Phe Gly lie Val Ser Gly Cys 105 110 Leu Ser Phe Pro Met Asp Asn Leu 115 120 Gin Gly Glu Leu Leu Asn Leu Tyr 125 Gin Val His Val Glu Asn Arg Val 130 135 Gly Ala Gin Arg Phe Arg Asp Ser 140 Arg Pro Trp Ser Glu Arg Pro Arg 145 150 Ala Val Ser Pro Glu Ala Ser Asp 155 160 Pro Arg Val Tyr Ser Asp Pro Ala 165 Ser Phe Val Ala Asn Gin Leu Gly 170 175 Leu Gly Pro Val Arg Tyr Ser Leu 180 Asp Ala Ala Cys Ala Ser Ala Leu 185 190 o Tyr Cys Leu Lys Leu Ala Ser Asp 195 200 His Leu Leu Ser Arg Ser Ala Asp 205 Val Met Leu Cys Gly Ala Thr Cys 210 215 Phe Pro Asp Pro Phe Phe lie Leu 220 Ser Gly Phe Ser Thr Phe Gin Ala _ 225 230 Met Pro Leu Gly Gly Pro Asp Asp 235 240 « Asn Pro Leu Ser Val Pro Leu Arg ^ 245 Gin Gly Ser Gin Gly Leu Thr Pro 250 255 Gly Glu Gly Gly Ala lie Met Val 260 Leu Lys Arg Leu Glu Asp Ala Val 265 270 Arg Asp Gly Asp Arg lie Tyr Gly 275 280 Thr Leu Leu Gly Thr Ser Leu Ser 285 —^ Asn Ala Gly Cys Gly Leu Pro Leu C* 290 295 Ser Pro His Leu Pro Ser Glu Lys 300 Ser Cys Met Glu Asp Leu Tyr Thr 305 310 Ser Val Gly lie Asp Pro Ser Glu 315 320 Val Gin Tyr Val Glu Cys His Ala 325 Thr Gly Thr Pro Gin Gly Asp Val 330 335 Val Glu Val Glu Ala Leu Arg His 340 Cys Phe Arg Gly Asn Thr Asp His 345 350 Pro Pro Arg Met Gly Ser Thr Lys 355 360 Gly Asn Phe Gly His Thr Leu Val 365 Ala Ala Gly Phe Ala Gly Met Ala 370 375 Lys Val Leu Leu Ser Met Gin His 380Ala Ser Thr Thr Ala Asn Leu Arg 100 Asp Phe Gly lie Val Ser Gly Cys 105 110 Leu Ser Phe Pro Met Asp Asn Leu 115 120 Gin Gly Glu Leu Leu Asn Leu Tyr 125 Gin Val His Val Glu Asn Arg Val 130 135 Gly Ala Gin Arg Phe Arg Asp Ser 140 Arg Pro Trp Ser Glu Arg Pro Arg 145 150 Ala Val Ser Pro Glu Ala Ser Asp 155 160 Pro Arg Val Tyr Ser Asp Pro Ala 165 Ser Phe Val Ala Asn Gin Leu Gly 170 175 Leu Gly Pro Val Arg Tyr Ser Leu 180 Asp Ala Ala Cys Ala Ser Ala Leu 185 190 o Tyr Cys Leu Lys Leu Ala Ser Asp 195 200 His Leu Leu Ser Arg Ser Ala Asp 205 Val Met Leu Cys Gly Ala Thr Cys 210 215 Phe Pro Asp Pro Phe Phe lie Leu 220 Ser Gly Phe Ser Thr Phe Gin Ala _ 225 230 Met Pro Leu Gly Gly Pro Asp Asp 235 240 « Asn Pro Leu Ser Val Pro Leu Arg ^ 245 Gin Gly Ser Gin Gly Leu Thr Pro 250 255 Gly Glu Gly Gly Ala lie Met Val 260 Leu Lys Arg Leu Glu Asp Ala Val 265 270 Arg Asp Gly Asp Arg lie Tyr Gly 275 280 Thr Leu Leu Gly Thr Ser Leu Ser 285 —^ Asn Ala Gly Cys Gly Leu Pro Leu C* 290 295 Ser Pro His Leu Pro Ser Glu Lys 300 Ser Cys Met Glu Asp Leu Tyr Thr 305 310 Ser Val Gly lie Asp Pro Ser Glu 315 320 Val Gin Tyr Val Glu Cys His Ala 325 Thr Gly Thr Pro Gin Gly Asp Val 330 335 Val Glu Val Glu Ala Leu Arg His 340 Cys Phe Arg Gly Asn Thr Asp His 345 350 Pro Pro Arg Met Gly Ser Thr Lys 355 360 Gly Asn Phe Gly His Thr Leu Val 365 Ala Ala Gly Phe Ala Gly Met Ala 370 375 Lys Val Leu Leu Ser Met Gin His 380

Gly Thr lie Pro Pro Thr Pro Gly Val Asp Arg Ser Asn Cys lie Asp 385 390 395 400 47 201038734Gly Thr lie Pro Pro Pro Pro Pro Gly Val Asp Arg Ser Asn Cys lie Asp 385 390 395 400 47 201038734

a 5 u u It—10 1 A 4 L G r r o g e e 3 r s s 4 A n r 1 e G sa 5 u u It—10 1 A 4 L G r r o g e e 3 r s s 4 A n r 1 e G s

Pro Leu Val Val Asp Glu Ala lie Pro Trp Pro Tyr Ser 405 410Pro Leu Val Val Asp Glu Ala lie Pro Trp Pro Tyr Ser 405 410

Ala Arg Ala Gly Lys Pro Gly Asp Glu Leu Lys Cys Ala 420 425Ala Arg Ala Gly Lys Pro Gly Asp Glu Leu Lys Cys Ala 420 425

Ala Phe Gly Phe Gly Gly Thr Asn Ala His Cys Val Phe 435 440 445 <210> 31 <211> 1293 <212> DNA <213> 裂殖壺菌物種 <400> 31 ggaccgattg ccatcatcgg gatggacgcg acgtttggta ccctcaaggg cctggacgcg 60 tttgagcagg ccatctacaa gggcacggac ggcgccagcg acctgccgag caagcgctgg 120 cggttcctgg gcgccgacac ggacttcttg accgccatgg gcctcgacgc cgtgccgcgc 180 gggtgctacg tgcgcgacgt ggacgtggac tacaagcggc tgcggtcgcc gatgatccct 240 gaggacgtcc tgcgcccgca acagctgctg gcggtggcta cgatggaccg cgcgctgcag 300 gacgctggaa tggcgacggg aggcaaggtg gcggtgctgg tggggctcgg cacggacacc 360 gagctgtacc ggcaccgcgc gcgcgtgaca ctcaaggagc ggctcgaccc ggccgcgttc 420 tcgcccgagc aggtgcagga gatgatggac tacatcaacg actgcggcac ctcgacgtcg 480 tacacgtcgt acatcggcaa cctcgtggcc acgcgcgtgt cctcgcagtg gggctttacg 540 ggcccgtcct tcaccgtcac cgaaggcgca aactcggtct accgctgcct cgagctgggc 600 aagttcctgc tcgacacgca ccaggtggac gccgtcgtgg tggccggcgt cgacctctgt 660 gccaccgccg agaaccttta cctcaaggcg cgccgctccg ccatcagccg acaggaccac 720 cctcgcgcca actttgaggc cagcgccgac gggtactttg ccggcgaggg cagcggcgcc 780 ctggtcctca agcgccaggc cgacgttggc tcagacgaca aggtctacgc cagtgtcgcg 840 ggcctcacgt gcgccgcgca gcccgctgaa gccgtgtcgc cgctactact ccaagtccac 900 aacgacgaca acgagaagag ggtggtggag atggtggagc tcgccgccga ctcgggtcgc 960 catgcgccgc acttggccaa ctcgccgctg agcgccgagt cgcagctgga gcaagtgtcc 1020 aagttgctcg cgcaccaggt gccgggctcg gtggccatcg gcagcgtgcg cgccaacgtg 1080 ggagacgtcg ggtacgcctc gggcgccgcg agcctcatca agacggcgct gtgcctccac 1140 aaccgctacc tcccggccaa cccgcagtgg gagcggccgg tggcgccggt ctccgaggcg 1200 ctgtttactt gcccgcgctc gcgtgcctgg ctgaagaacc cgggcgagtc gcgactggcg 1260 gctgtcgcca gtgcctccga gagcgggtcc tgc 1293 <210> 32 <211> 431 <212> PRT <213〉 裂殖壺菌物種 <400> 32Ala Phe Gly Phe Gly Gly Thr Asn Ala His Cys Val Phe 435 440 445 <210> 31 <211> 1293 <212> DNA <213> Schizochytrium species <400> 31 ggaccgattg ccatcatcgg gatggacgcg acgtttggta ccctcaaggg cctggacgcg 60 tttgagcagg ccatctacaa gggcacggac ggcgccagcg acctgccgag caagcgctgg 120 cggttcctgg gcgccgacac ggacttcttg accgccatgg gcctcgacgc cgtgccgcgc 180 gggtgctacg tgcgcgacgt ggacgtggac tacaagcggc tgcggtcgcc gatgatccct 240 gaggacgtcc tgcgcccgca acagctgctg gcggtggcta cgatggaccg cgcgctgcag 300 gacgctggaa tggcgacggg aggcaaggtg gcggtgctgg tggggctcgg cacggacacc 360 gagctgtacc ggcaccgcgc gcgcgtgaca ctcaaggagc ggctcgaccc ggccgcgttc 420 tcgcccgagc aggtgcagga gatgatggac tacatcaacg actgcggcac ctcgacgtcg 480 tacacgtcgt acatcggcaa cctcgtggcc acgcgcgtgt cctcgcagtg gggctttacg 540 ggcccgtcct tcaccgtcac cgaaggcgca aactcggtct accgctgcct cgagctgggc 600 aagttcctgc tcgacacgca ccaggtggac gccgtcgtgg tggccggcgt cgacctctgt 660 gccaccgccg agaaccttta cctcaaggcg cgccgctccg ccatcagccg a caggaccac 720 cctcgcgcca actttgaggc cagcgccgac gggtactttg ccggcgaggg cagcggcgcc 780 ctggtcctca agcgccaggc cgacgttggc tcagacgaca aggtctacgc cagtgtcgcg 840 ggcctcacgt gcgccgcgca gcccgctgaa gccgtgtcgc cgctactact ccaagtccac 900 aacgacgaca acgagaagag ggtggtggag atggtggagc tcgccgccga ctcgggtcgc 960 catgcgccgc acttggccaa ggtacgcctc gggcgccgcg agcctcatca ctcgccgctg agcgccgagt cgcagctgga gcaagtgtcc 1020 aagttgctcg cgcaccaggt gccgggctcg gtggccatcg gcagcgtgcg cgccaacgtg 1080 ggagacgtcg agacggcgct gtgcctccac 1140 aaccgctacc tcccggccaa cccgcagtgg gagcggccgg tggcgccggt ctccgaggcg 1200 ctgtttactt gcccgcgctc gcgtgcctgg ctgaagaacc cgggcgagtc gcgactggcg 1260 gctgtcgcca gtgcctccga gagcgggtcc tgc 1293 < 210 > 32 < 211 > 431 < 212 > PRT < 213> Schizochytrium species < 400 > 32

Gly Pro 工le Ma lie lie Gly Met Asp Ala Thr Phe Gly Thr Leu Lys 15 10 15 48 201038734Gly Pro work le lie lie Gly Met Asp Ala Thr Phe Gly Thr Leu Lys 15 10 15 48 201038734

Gly Leu Asp Ala Phe Glu Gin Ala lie Tyr Lys Gly Thr Asp Gly Ala 20 25 30Gly Leu Asp Ala Phe Glu Gin Ala lie Tyr Lys Gly Thr Asp Gly Ala 20 25 30

Ser Asp Leu Pro Ser Lys Arg Trp Arg Phe Leu Gly Ala Asp Thr Asp 35 40 45Ser Asp Leu Pro Ser Lys Arg Trp Arg Phe Leu Gly Ala Asp Thr Asp 35 40 45

Phe Leu Thr Ala Met Gly Leu Asp Ala Val Pro Arg Gly Cys Tyr Val 50 55 60Phe Leu Thr Ala Met Gly Leu Asp Ala Val Pro Arg Gly Cys Tyr Val 50 55 60

Arg Asp Val Asp Val Asp Tyr Lys Arg Leu Arg Ser Pro Met 工le Pro 65 70 75 80Arg Asp Val Asp Val Asp Tyr Lys Arg Leu Arg Ser Pro Met Le Pro 65 70 75 80

Glu Asp Val Leu Arg Pro Gin Gin Leu Leu Ala Val Ala Thr Met Asp 85 90 95Glu Asp Val Leu Arg Pro Gin Gin Leu Leu Ala Val Ala Thr Met Asp 85 90 95

Arg Ala Leu Gin Asp Ala Gly Met Ala Thr Gly Gly Lys Val Ala Val 100 105 110Arg Ala Leu Gin Asp Ala Gly Met Ala Thr Gly Gly Lys Val Ala Val 100 105 110

Leu Val Gly Leu Gly Thr Asp Thr Glu Leu Tyr Arg His Arg Ala Arg 115 120 125Leu Val Gly Leu Gly Thr Asp Thr Glu Leu Tyr Arg His Arg Ala Arg 115 120 125

Val Thr Leu Lys Glu Arg Leu Asp Pro Ala Ala Phe Ser Pro Glu Gin 130 135 140Val Thr Leu Lys Glu Arg Leu Asp Pro Ala Ala Phe Ser Pro Glu Gin 130 135 140

Val Gin Glu Met Met Asp Tyr 工le Asn Asp Cys Gly Thr Ser Thr Ser 145 150 155 160Val Gin Glu Met Met Asp Tyr Labor As As As As Cy Cys Gly Thr Ser Thr Ser 145 150 155 160

Tyr Thr Ser Tyr lie Gly Asn Leu Val Ala Thr Arg Val Ser Ser Gin 165 170 175Tyr Thr Ser Tyr lie Gly Asn Leu Val Ala Thr Arg Val Ser Ser Gin 165 170 175

Trp Gly Phe Thr Gly Pro Ser Phe Thr Val Thr Glu Gly Ala Asn Ser 180 185 190Trp Gly Phe Thr Gly Pro Ser Phe Thr Val Thr Glu Gly Ala Asn Ser 180 185 190

Val Tyr Arg Cys Leu Glu Leu Gly Lys Phe Leu Leu Asp Thr His Gin 195 200 205Val Tyr Arg Cys Leu Glu Leu Gly Lys Phe Leu Leu Asp Thr His Gin 195 200 205

Val Asp Ala Val Val Val Ala Gly Val Asp Leu Cys Ala Thr Ala Glu 210 215 220Val Asp Ala Val Val Val Ala Gly Val Asp Leu Cys Ala Thr Ala Glu 210 215 220

Asn Leu Tyr Leu Lys Ala Arg Arg Ser Ala lie Ser Arg Gin Asp His 225 230 235 240Asn Leu Tyr Leu Lys Ala Arg Arg Ser Ala lie Ser Arg Gin Asp His 225 230 235 240

Pro Arg Ala Asn Phe Glu Ala Ser Ala Asp Gly Tyr Phe Ala Gly Glu 245 250 255Pro Arg Ala Asn Phe Glu Ala Ser Ala Asp Gly Tyr Phe Ala Gly Glu 245 250 255

Gly Ser Gly Ala Leu Val Leu Lys Arg Gin Ala Asp Val Gly Ser Asp 260 265 270Gly Ser Gly Ala Leu Val Leu Lys Arg Gin Ala Asp Val Gly Ser Asp 260 265 270

Asp Lys Val Tyr Ala Ser Val Ala Gly Leu Thr Cys Ala Ala Gin Pro 275 280 285Asp Lys Val Tyr Ala Ser Val Ala Gly Leu Thr Cys Ala Ala Gin Pro 275 280 285

Ala Glu Ala Val Ser Pro Leu Leu Leu Gin Val His Asn Asp Asp Asn 290 295 300Ala Glu Ala Val Ser Pro Leu Leu Leu Gin Val His Asn Asp Asp Asn 290 295 300

Glu Lys Arg Val Val Glu Met Val Glu Leu Ala Ala Asp Ser Gly Arg 305 310 315 320 49 201038734Glu Lys Arg Val Val Glu Met Val Glu Leu Ala Ala Asp Ser Gly Arg 305 310 315 320 49 201038734

His Ala Pro His Leu Ala Asn Ser Pro Leu Ser Ala Glu Ser Gin Leu 325 330 335His Ala Pro His Leu Ala Asn Ser Pro Leu Ser Ala Glu Ser Gin Leu 325 330 335

Glu Gin Val Ser Lys Leu Leu Ala His Gin Val Pro Gly Ser Val Ala 340 345 350 lie Gly Ser Val Arg Ala Asn Val Gly Asp Val Gly Tyr Ala Ser Gly 355 360 365Glu Gin Val Ser Lys Leu Leu Ala His Gin Val Pro Gly Ser Val Ala 340 345 350 lie Gly Ser Val Arg Ala Asn Val Gly Asp Val Gly Tyr Ala Ser Gly 355 360 365

Ala Ala Ser Leu lie Lys Thr Ala Leu Cys Leu His Asn Arg Tyr Leu 370 375 380 a _—_ A o 5 Γ 8 p 3Ala Ala Ser Leu lie Lys Thr Ala Leu Cys Leu His Asn Arg Tyr Leu 370 375 380 a ___ A o 5 Γ 8 p 3

Asn Pro Gin Trp Glu Arg Pro Val Ala Pro Val Ser Glu Ala 390 395 400Asn Pro Gin Trp Glu Arg Pro Val Ala Pro Val Ser Glu Ala 390 395 400

Leu Phe Thr Cys Pro Arg Ser Arg Ala Trp Leu Lys Asn Pro Gly Glu 〇 405 410 415Leu Phe Thr Cys Pro Arg Ser Arg Ala Trp Leu Lys Asn Pro Gly Glu 〇 405 410 415

Ser Arg Leu Ala Ala Val Ala Ser Ala Ser Glu Ser Gly Ser Cys 420 425 430 <210> 33 <211> 1203 <212> DMA <213〉 裂i壺菌物種 <4〇〇> 33 acgccggaga agctggagaa ggagttggag ctggcagcca agggtgtacc gcgaagcgcc 60 aaggccgggc gcaactggat gtcgccatcg ggcagcgcct ttgcgccgac acctgtgacc 120 agcgaccgcg tcgcgttcat gtacggcgag ggccgcagcc cctactacgg cgtcgggctc 180 gacctgcacc gcctgtggcc ggctttgcac gagcgcatca acgacaagac cgcggcgctg 240 tgggagaacg gcgactcgtg gctcatgccg cgcgcggtgg atgccgactc gcagcgcgcc 300 gtgcagacgg cctttgacgc ggaccagatc gagatgttcc gcacgggcat cttcgtgtcc 360 atctgcctca ccgactacgc gcgcgacgtg ctcggggtgc agcccaaggc gtgcttcggc 420 i ctcagcctcg gcgagatctc catgctcttt gcgctgtcgc gacgcaactg cggcctgtcg 480 gaccagctca cgcagcgcct acgcacctcg ccggtgtggt cgacacagct ggcggtggag 540 ttccaggcct tgcgcaagct atggaacgtg ccggcggacg cccccgtgga gtccttctgg 600 cagggctact tggttcgcgc cagccgcgcc gaaatcgaga aggcgatcgg gcccgacaac 660 cgcttcgtgc gcctgctgat cgtcaacgac tcgagcagcg cgctgatcgc cggcaaacct 720 gccgagtgtc tgcgcgtgct ggagcgcctg ggcgggcggt tgccgccgat gcccgtcaag 780 caaggcatga ttgggcactg ccccgaagtg gcgccctaca cgccgggcat cgcgcacatc 840 cacgagattt tggagattcc ggacagcccc gtcaagatgt acacctcggt caccaacgcc 900 gagctgcgcg ggggcagcaa cagcagcatc accgagttcg tgcagaagtt gtacacgcgc 960 atcgccgact ttccgggcat cgtcgacaag gtcagccgtg acggccacga tgtcttcgtc 1020 gaggtggggc cgaacaacat gcgctccgcc gcggtcagtg acattcttgg caaggctgcc 1080 accccgcatg tctccgtggc gctggaccgc cccagtgagt cggcgtggac gcagaccctc 1140 aagtcgctgg cgctgctgac cgcccaccgc gtgcccctgc acaacccgac tctgtttgcg 1200 50 1203 201038734 gac <210> 34 <211> 401 <212> PRT <213> 裂殖壺菌物種 <400> 34Ser Arg Leu Ala Ala Val Ala Ser Ala Ser Glu Ser Gly Ser Cys 420 425 430 <210> 33 <211> 1203 <212> DMA <213> Schizophrenia species <4〇〇> 33 acgccggaga agctggagaa ggagttggag ctggcagcca agggtgtacc gcgaagcgcc 60 aaggccgggc gcaactggat gtcgccatcg ggcagcgcct ttgcgccgac acctgtgacc 120 agcgaccgcg tcgcgttcat gtacggcgag ggccgcagcc cctactacgg cgtcgggctc 180 gacctgcacc gcctgtggcc ggctttgcac gagcgcatca acgacaagac cgcggcgctg 240 tgggagaacg gcgactcgtg gctcatgccg cgcgcggtgg atgccgactc gcagcgcgcc 300 gtgcagacgg cctttgacgc ggaccagatc gagatgttcc gcacgggcat cttcgtgtcc 360 atctgcctca ccgactacgc gcgcgacgtg ctcggggtgc agcccaaggc gtgcttcggc 420 i ctcagcctcg gcgagatctc catgctcttt gcgctgtcgc gacgcaactg cggcctgtcg 480 gaccagctca cgcagcgcct acgcacctcg ccggtgtggt cgacacagct ggcggtggag 540 ttccaggcct tgcgcaagct atggaacgtg ccggcggacg cccccgtgga gtccttctgg 600 cagggctact tggttcgcgc cagccgcgcc gaaatcgaga aggcgatcgg gcccgacaac 660 cgcttcgtgc gcctgctgat cgtcaacgac tcgagcagcg cgctgatcgc cggcaaa cct 720 gccgagtgtc tgcgcgtgct ggagcgcctg ggcgggcggt tgccgccgat gcccgtcaag 780 caaggcatga ttgggcactg ccccgaagtg gcgccctaca cgccgggcat cgcgcacatc 840 cacgagattt tggagattcc ggacagcccc gtcaagatgt acacctcggt caccaacgcc 900 gagctgcgcg ggggcagcaa cagcagcatc accgagttcg tgcagaagtt gtacacgcgc 960 atcgccgact ttccgggcat cgtcgacaag gtcagccgtg acggccacga tgtcttcgtc 1020 gaggtggggc cgaacaacat gcgctccgcc gcggtcagtg acattcttgg caaggctgcc 1080 accccgcatg tctccgtggc gctggaccgc cccagtgagt cggcgtggac gcagaccctc 1140 aagtcgctgg cgctgctgac cgcccaccgc gtgcccctgc acaacccgac tctgtttgcg 1200 50 1203 201038734 gac <210> 34 <211> 401 <212> PRT <213> Schizochytrium species <400>

Thr Pro Glu Lys Leu Glu Lys Glu Leu Glu Leu Ala Ala Lys Gly Val 15 10 15Thr Pro Glu Lys Leu Glu Lys Glu Leu Glu Leu Ala Ala Lys Gly Val 15 10 15

Pro Arg Ser Ala Lys Ala Gly Arg Asn Trp Met Ser Pro Ser Gly Ser 20 25 30Pro Arg Ser Ala Lys Ala Gly Arg Asn Trp Met Ser Pro Ser Gly Ser 20 25 30

Ala Phe Ala Pro Thr Pro Val Thr Ser Asp Arg Val Ala Phe Met Tyr 35 40 45 〇 〇Ala Phe Ala Pro Thr Pro Val Thr Ser Asp Arg Val Ala Phe Met Tyr 35 40 45 〇 〇

Gly Glu Gly Arg Ser Pro Tyr Tyr Gly Val Gly Leu Asp Leu His Arg 50 55 60Gly Glu Gly Arg Ser Pro Tyr Tyr Gly Val Gly Leu Asp Leu His Arg 50 55 60

Leu Trp Pro Ala Leu His Glu Arg 工le Asn Asp Lys Thr Ala Ala Leu 65 70 75 80Leu Trp Pro Ala Leu His Glu Arg Le Asn Asp Lys Thr Ala Ala Leu 65 70 75 80

Trp Glu Asn Gly Asp Ser Trp Leu Met Pro Arg Ala Val Asp Ala Asp 85 90 95Trp Glu Asn Gly Asp Ser Trp Leu Met Pro Arg Ala Val Asp Ala Asp 85 90 95

Ser Gin Arg Ala Val Gin Thr Ala Phe Asp Ala Asp Gin lie Glu Met 100 105 110Ser Gin Arg Ala Val Gin Thr Ala Phe Asp Ala Asp Gin lie Glu Met 100 105 110

Phe Arg Thr Gly lie Phe Val Ser lie Cys Leu Thr Asp Tyr Ala Arg 115 120 125Phe Arg Thr Gly lie Phe Val Ser lie Cys Leu Thr Asp Tyr Ala Arg 115 120 125

Asp Val Leu Gly Val Gin Pro Lys Ala Cys Phe Gly Leu Ser Leu Gly 130 135 140Asp Val Leu Gly Val Gin Pro Lys Ala Cys Phe Gly Leu Ser Leu Gly 130 135 140

Glu lie Ser Met Leu Phe Ala Leu Ser Arg Arg Asn Cys Gly Leu Ser 145 150 155 160Glu lie Ser Met Leu Phe Ala Leu Ser Arg Arg Asn Cys Gly Leu Ser 145 150 155 160

Asp Gin Leu Thr Gin Arg Leu Arg Thr Ser Pro Val Trp Ser Thr Gin 165 170 175Asp Gin Leu Thr Gin Arg Leu Arg Thr Ser Pro Val Trp Ser Thr Gin 165 170 175

Leu Ala Val Glu Phe Gin Ala Leu Arg Lys Leu Trp Asn Val Pro Ala 180 185 190Leu Ala Val Glu Phe Gin Ala Leu Arg Lys Leu Trp Asn Val Pro Ala 180 185 190

Asp Ala Pro Val Glu Ser Phe Trp Gin Gly Tyr Leu Val Arq Ala Ser 195 200 205Asp Ala Pro Val Glu Ser Phe Trp Gin Gly Tyr Leu Val Arq Ala Ser 195 200 205

Arg Ala Glu lie Glu Lys Ala lie Gly Pro Asp Asn Arg Phe Val Arg 210 215 220Arg Ala Glu lie Glu Lys Ala lie Gly Pro Asp Asn Arg Phe Val Arg 210 215 220

Leu Leu lie Val Asn Asp Ser Ser Ser Ala Leu lie Ala Gly Lys Pro 225 230 235 240Leu Leu lie Val Asn Asp Ser Ser Ser Ala Leu lie Ala Gly Lys Pro 225 230 235 240

Ala Glu Cys Leu Arg Val Leu Glu Arg Leu Gly Gly Arg Leu 245 250 0 5 r 5 p 2 o r p 51 201038734Ála Glu Arg Leu Gly Arg Leu 245 250 0 5 r 5 p 2 o r p 51 201038734

Met Pro Val Lys Gin Gly Met lie Gly His Cys Pro Glu Val Ala Pro 260 265 270Met Pro Val Lys Gin Gly Met lie Gly His Cys Pro Glu Val Ala Pro 260 265 270

Tyr Thr Pro Gly lie Ala His lie His Glu lie Leu Glu lie Pro Asp 275 280 285Tyr Thr Pro Gly lie Ala His lie His Glu lie Leu Glu lie Pro Asp 275 280 285

Ser Pro Val Lys Met Tyr Thr Ser Val Thr Asn Ala Glu Leu Arg Gly 290 295 300Ser Pro Val Lys Met Tyr Thr Ser Val Thr Asn Ala Glu Leu Arg Gly 290 295 300

Gly Ser Asn Ser Ser lie Thr Glu Phe Val Gin Lys Leu Tyr Thr Arg 305 310 315 320 lie Ala Asp Phe Pro Gly lie Val Asp Lys Val Ser Arg Asp Gly His 325 330 335Gly Ser Asn Ser Ser lie Thr Glu Phe Val Gin Lys Leu Tyr Thr Arg 305 310 315 320 lie Ala Asp Phe Pro Gly lie Val Asp Lys Val Ser Arg Asp Gly His 325 330 335

Asp Val Phe Val Glu Val Gly Pro Asn Asn Met Arg Ser Ala Ala Val 340 345 350Asp Val Phe Val Glu Val Gly Pro Asn Asn Met Arg Ser Ala Ala Val 340 345 350

Ser Asp lie Leu Gly Lys Ala Ala Thr Pro His Val Ser Val Ala Leu 355 360 365Ser Asp lie Leu Gly Lys Ala Ala Thr Pro His Val Ser Val Ala Leu 355 360 365

Asp Arg Pro Ser Glu Ser Ala Trp Thr Gin Thr Leu Lys Ser Leu Ala 370 3Ί5 380Asp Arg Pro Ser Glu Ser Ala Trp Thr Gin Thr Leu Lys Ser Leu Ala 370 3Ί5 380

Leu Leu Thr Ala His Arg Val Pro Leu His Asn Pro Thr Leu Phe Ala 385 390 395 400Leu Leu Thr Ala His Arg Val Pro Leu His Asn Pro Thr Leu Phe Ala 385 390 395 400

Asp <210> 35 <211> 1273 <212> DNA <213> 裂殖壺菌物種 <400> 35 tacgacgtgg actggccgct ctacatgggc gccatggcgg aaggcatctc gtcggtagac 60 ctggtggtcg ctgccgccga ggcccgcatg ctggcatcat tcggagcggc ccgcttgcct 120Asp <210> 35 <211> 1273 <212> DNA <213> Schizochytrium species <400> 35 tacgacgtgg actggccgct ctacatgggc gccatggcgg aaggcatctc gtcggtagac 60 ctggtggtcg ctgccgccga ggcccgcatg ctggcatcat tcggagcggc ccgcttgcct 120

atggaccagg tggaactcca gatccgtgag atccagcaac gcacctccaa cgcctttgct 180 gtcaacctga tgccgggtcc tgacgaggcc gcgacggtgg acgcgctgct gcgcacgggc 240 gtctcaatcg tcgaggcatc gggctacacc ggcgcgctct ctgcagacct ggtgcgctac 300 cgtgtcacgg gtctgcgacg aactagttgc ggtgcttctg tgtcggcgac tcaccgtgtg 360 gtcgccaagg tgtcgcgcac cgaggtggcc gagcactttc tgcgcccggc gccggccgcc 420 gtactagagg ctttggtcgc cgccaaacag attacgcccg agcaggccgc gctggccagc 480 cgcgtcgcca tggccgacga cgtcgcggtg gaggccgact cgggcgggca caccgacaac 540 cgaccgatcc acgtgctgct gccgctcgtg gtggcgcagc gcaaccgctg gcgccacctg 600 gtggacacgc cagtgcgcgt cggcgccggc ggcgggatcg cctgtccgcg cgccgcgctg 660 ctcgcctttt ccctgggcgc cgcctttgtg gtcaccgggt ccgtcaacca actggcccgc 720 gaggctggca ccagcgacgc ggtccgacta ctgctggcga cggccaccta ctcggacgtg 780 gccatggcgc cgggcggcgt ccaggtgctc aagaagcaga ccatgttcgc cgcgcgggcc 840 acgatgctcg cccagctgca ggccaagttc ggctcctttg acgccgtgcc ggagccgcag 900 52 960 201038734 ctgcgcaagc tcgagcgctc cgtgttcaag cagtccgtgg cggacgtgtg ggctgctgca cgcgaaaagt ttggtgtcga cgctaccgct gcaagtccgc aggagaggat ggcgctctgt gtgcgctggt acatgtcgca gtcgtcgcga tgggctaccg aggcgacgtc cgcgcgcaag gcggactacc agatctggtg cggccccgcc atcggcagct tcaacgactt cgttcgcggc accaagctgg acgcgaccgc tggcaccggc gagtttccgc gcgtcgtgga catcaaccag cacatcctcc tcggagcctc gcactaccgc cgcgtgcagc aacaacaaca ggacgacgac gtagaataca tea <210> 36 <211> 401 <212> PRT <213> 裂殖壺菌物種 <400> 36atggaccagg tggaactcca gatccgtgag atccagcaac gcacctccaa cgcctttgct 180 gtcaacctga tgccgggtcc tgacgaggcc gcgacggtgg acgcgctgct gcgcacgggc 240 gtctcaatcg tcgaggcatc gggctacacc ggcgcgctct ctgcagacct ggtgcgctac 300 cgtgtcacgg gtctgcgacg aactagttgc ggtgcttctg tgtcggcgac tcaccgtgtg 360 gtcgccaagg tgtcgcgcac cgaggtggcc gagcactttc tgcgcccggc gccggccgcc 420 gtactagagg ctttggtcgc cgccaaacag attacgcccg agcaggccgc gctggccagc 480 cgcgtcgcca tggccgacga cgtcgcggtg gaggccgact cgggcgggca caccgacaac 540 cgaccgatcc acgtgctgct gccgctcgtg gtggcgcagc gcaaccgctg gcgccacctg 600 gtggacacgc cagtgcgcgt cggcgccggc ggcgggatcg cctgtccgcg cgccgcgctg 660 ctcgcctttt ccctgggcgc cgcctttgtg gtcaccgggt ccgtcaacca actggcccgc 720 gaggctggca ccagcgacgc ggtccgacta ctgctggcga cggccaccta ctcggacgtg 780 gccatggcgc cgggcggcgt ccaggtgctc aagaagcaga ccatgttcgc cgcgcgggcc 840 acgatgctcg cccagctgca ggccaagttc ggctcctttg acgccgtgcc ggagccgcag 900 52 960 201038734 ctgcgcaagc tcgagcgctc cgtgttcaag cagtccgtgg cggacgtgtg ggctgctgca Cgcgaaa agt ttggtgtcga cgctaccgct gcaagtccgc aggagaggat ggcgctctgt gtgcgctggt acatgtcgca gtcgtcgcga tgggctaccg aggcgacgtc cgcgcgcaag gcggactacc agatctggtg cggccccgcc atcggcagct tcaacgactt cgttcgcggc accaagctgg acgcgaccgc tggcaccggc gagtttccgc gcgtcgtgga catcaaccag cacatcctcc tcggagcctc gcactaccgc cgcgtgcagc aacaacaaca ggacgacgac gtagaataca tea < 210 > 36 < 211 > 401 < 212 > PRT < 213 > Schizochytrium species <400> 36

Tyr Asp Val Asp Trp Pro Leu Tyr Met Gly Ala Met Ala Glu Gly lie 1020 1080 1140 1200 1260 1273Tyr Asp Val Asp Trp Pro Leu Tyr Met Gly Ala Met Ala Glu Gly lie 1020 1080 1140 1200 1260 1273

Ser Ser Val Asp Leu Val Val Ala Ala Ala Glu Ala Arg Met Leu Ala 20 25 30Ser Ser Val Asp Leu Val Val Ala Ala Ala Glu Ala Arg Met Leu Ala 20 25 30

Ser Phe Gly Ala Ala Arg Leu Pro Met Asp Gin Val Glu Leu Gin lie 35 40 45Ser Phe Gly Ala Ala Arg Leu Pro Met Asp Gin Val Glu Leu Gin lie 35 40 45

Arg Glu 工le Gin Gin Arg Thr Ser Asn Ala Phe Ala Val Asn Leu Met 50 55 60Arg Glu gong Gin Gin Arg Thr Ser Asn Ala Phe Ala Val Asn Leu Met 50 55 60

Pro Gly Pro Asp Glu Ala Ala Thr Val Asp Ala Leu Leu Arg Thr Gly 65 70 75 80Pro Gly Pro Asp Glu Ala Ala Thr Val Asp Ala Leu Leu Arg Thr Gly 65 70 75 80

Val Ser lie Val Glu Ala Ser Gly Tyr Thr Gly Ala Leu Ser Ala Asp 85 90 95Val Ser lie Val Glu Ala Ser Gly Tyr Thr Gly Ala Leu Ser Ala Asp 85 90 95

Leu Val Arg Tyr Arg Val Thr Gly Leu Arg Arg Thr Ser Cys Gly Ala 100 105 110Leu Val Arg Tyr Arg Val Thr Gly Leu Arg Arg Thr Ser Cys Gly Ala 100 105 110

Ser Val Ser Ala Thr His Arg Val Val Ala Lys Val Ser Arg Thr Glu 115 120 125Ser Val Ser Ala Thr His Arg Val Val Ala Lys Val Ser Arg Thr Glu 115 120 125

Val Ala Glu His Phe Leu Arg Pro Ala Pro Ala Ala Val Leu Glu Ala 130 135 140Val Ala Glu His Phe Leu Arg Pro Ala Pro Ala Ala Val Leu Glu Ala 130 135 140

Leu Val Ala Ala Lys Gin lie Thr Pro Glu Gin Ala Ala Leu Ala Ser 145 150 155 160Leu Val Ala Ala Lys Gin lie Thr Pro Glu Gin Ala Ala Leu Ala Ser 145 150 155 160

Arg Val Ala Met Ala Asp Asp Val Ala Val Glu Ala Asp Ser Gly Gly 165 170 175Arg Val Ala Met Ala Asp Asp Val Ala Val Glu Ala Asp Ser Gly Gly 165 170 175

His Thr Asp Asn Arg Pro lie His Val Leu Leu Pro Leu Val Val Ala 180 185 190His Thr Asp Asn Arg Pro lie His Val Leu Leu Pro Leu Val Val Ala 180 185 190

Gin Arg Asn Arg Trp Arg His Leu Val Asp Thr Pro Val Arq Val Gly 195 200 205Gin Arg Asn Arg Trp Arg His Leu Val Asp Thr Pro Val Arq Val Gly 195 200 205

Ala Gly Gly Gly lie Ala Cys Pro Arg Ala Ala Leu Leu Ala Phe Ser 53 201038734 210 215 220Ala Gly Gly Gly lie Ala Cys Pro Arg Ala Ala Leu Leu Ala Phe Ser 53 201038734 210 215 220

Leu Gly Ala Ala Phe Val Val Thr Gly Ser Val Asn Gin Leu Ala Arg 225 230 235 240Leu Gly Ala Ala Phe Val Val Thr Gly Ser Val Asn Gin Leu Ala Arg 225 230 235 240

Glu Ala Gly Thr Ser Asp Ala Val Arg Leu Leu Leu Ala Thr Ala Thr 245 250 255Glu Ala Gly Thr Ser Asp Ala Val Arg Leu Leu Leu Ala Thr Ala Thr 245 250 255

Tyr Ser Asp Val Ala Met Ala Pro Gly Gly Val Gin Val Leu Lys Lys 260 265 270Tyr Ser Asp Val Ala Met Ala Pro Gly Gly Val Gin Val Leu Lys Lys 260 265 270

GG

t 5 e 7 M 2 r h Tt 5 e 7 M 2 r h T

Phe Ala Ala Arg Ala Thr Met Leu Ala Gin Leu Gin Ala 280 285Phe Ala Ala Arg Ala Thr Met Leu Ala Gin Leu Gin Ala 280 285

Lys Phe Glv Ser Phe Asp Ala Val Pro Glu Pro Gin Leu Arg Lys Leu 290 295 300Lys Phe Glv Ser Phe Asp Ala Val Pro Glu Pro Gin Leu Arg Lys Leu 290 295 300

Glu Arg Ser Val Phe Lys Gin Ser Val Ala Asp Val Trp Ala Ala Ala 305 310 315 320Glu Arg Ser Val Phe Lys Gin Ser Val Ala Asp Val Trp Ala Ala Ala 305 310 315 320

Arq Glu Lys Phe Gly Val Asp Ala Thr Ala Ala Ser Pro Gin Glu Arg 325 330 335Arq Glu Lys Phe Gly Val Asp Ala Thr Ala Ala Ser Pro Gin Glu Arg 325 330 335

Met Ala Leu Cys Val Arg Trp Tyr Met Ser Gin Ser Ser Arg Trp Ala 340 345 350Met Ala Leu Cys Val Arg Trp Tyr Met Ser Gin Ser Ser Arg Trp Ala 340 345 350

Thr Glu Ala Thr Ser Ala Arg Lys Ala Asp Tyr Gin lie Trp Cys Gly 355 360 365Thr Glu Ala Thr Ser Ala Arg Lys Ala Asp Tyr Gin lie Trp Cys Gly 355 360 365

Pro Ala lie Gly Ser Phe Asn Asp Phe Val Arg Gly Thr Lys Leu Asp 370 375 380Pro Ala lie Gly Ser Phe Asn Asp Phe Val Arg Gly Thr Lys Leu Asp 370 375 380

Ala Thr Ala Gly Thr Gly Glu Phe Pro Arg Val Val Asp lie Asn Gin 385 390 395 400Ala Thr Ala Gly Thr Gly Glu Phe Pro Arg Val Val Asp lie Asn Gin 385 390 395 400

His <210> 37 <211> 1350 <212> DNA <213〉 裂殖壺菌物種 <400> 37 atgacatcat cgaagaagac tcccgtgtgg gagatgagca aggaggagct gctggacggc 60 aagacggtgg tcttcgacta caacgagctg ctcgaattcg ccgagggcga cgtgggccaa 120 gtgttcggac ccgagttcga catcatcgac aagtaccggc gtcgcgtgcg gctgccggcg 180 cgcgagtacc tgctcgtgtc gcgcgtgacg ctgatggacg ccgaggtgaa caacttccgc 240 gtcgggtcgc gcatggtgac cgagtacgac gtgcccgtga acggggagct gtcggagggc 300 ggggacgtgc cgtgggcggt gctggtggag tcggggcagt gcgacctgat gctcatctcg 360 tacatgggca tcgacttcca gtgcaagggc gaccgcgtgt accgcctgct caacacatcg 420 ctcaccttct tcggggtggc gcacgagggc gagacgctgg tgtacgacat ccgcgtcacg 480 gggttcgcca agggcgcggg cggggagatc tcgatgttct tcttcgagta cgactgcttc 540 gtggacggcc gcctgctgat cgagatgcgc gacgggtgcg ccgggttctt cacggacgcc 600 54 660 201038734His < 210 > 37 < 211 > 1350 < 212 > DNA < 213> Schizochytrium species < 400 > 37 atgacatcat cgaagaagac tcccgtgtgg gagatgagca aggaggagct gctggacggc 60 aagacggtgg tcttcgacta caacgagctg ctcgaattcg ccgagggcga cgtgggccaa 120 gtgttcggac ccgagttcga catcatcgac aagtaccggc gtcgcgtgcg gctgccggcg 180 cgcgagtacc tgctcgtgtc gcgcgtgacg ctgatggacg ccgaggtgaa caacttccgc 240 gtcgggtcgc gcatggtgac cgagtacgac gtgcccgtga acggggagct gtcggagggc 300 ggggacgtgc cgtgggcggt gctggtggag tcggggcagt gcgacctgat gctcatctcg 360 tacatgggca tcgacttcca gtgcaagggc gaccgcgtgt accgcctgct caacacatcg 420 ctcaccttct tcggggtggc gcacgagggc gagacgctgg tgtacgacat ccgcgtcacg 480 gggttcgcca agggcgcggg cggggagatc tcgatgttct tcttcgagta cgactgcttc 540 gtggacggcc gcctgctgat cgagatgcgc gacgggtgcg ccgggttctt cacggacgcc 600 54 660 201038734

720 780 840 900 960 1020 1080 1140 1200 1260 1320 1350 gagctggccg ccggcaaggg cgtgcttaag accaaggcgg agctggcggc gcgcgcgcag atccagaagc aggacatcgc gccctttgcg ccggcgccgt gctcgcacaa gacctcgctg gacgcgcgcg agatgcggct gctcgtggac cgccagtggg cgcgcgtctt cggcagcggc atggcgggca tcgactacaa gttgtgcgct cgcaagatgc tcatgatcga ccgcgtcacg cacctcgacc cgcgcggcgg cgcgcacggc ctcgggctgc tgatcgggga gaaggtgctg gagcgcgacc actggtactt cccctgccac tttgtgcgcg acgaggtgat ggccgggtcg ctggtcagcg acggctgctc gcagctcctc aaggtgtaca tgctgtggct cggcctgcac acgaccgtgg gcgcgttcga ctttcgtccc gtgagcgggc acgccaacaa ggtgcggtgc cgcgggcaga tctcaccgca caagggcaag ctcgtgtacg tgatggagat caaggaaatg ggctttgacg cgaagacggg cgatccgttt gcgatcgcgg acgtggacat catcgacgtc aacttcgagg agggacaggc gtttgcggga gtggaagacc tgcacagcta cggccagggc gacctccgca agaagatcgt cgtcgacttc aagggcatcg cgctctccct gcagaagcgg aaggagcagc agaaggaaag catgaccgtg <210> 38 <211> 450 <212> PRT <213〉 裂殖壺菌物種 <400> 38720 780 840 900 960 1020 1080 1140 1200 1260 1320 1350 gagctggccg ccggcaaggg cgtgcttaag accaaggcgg agctggcggc gcgcgcgcag atccagaagc aggacatcgc gccctttgcg ccggcgccgt gctcgcacaa gacctcgctg gacgcgcgcg agatgcggct gctcgtggac cgccagtggg cgcgcgtctt cggcagcggc atggcgggca tcgactacaa gttgtgcgct cgcaagatgc tcatgatcga ccgcgtcacg cacctcgacc cgcgcggcgg cgcgcacggc ctcgggctgc tgatcgggga gaaggtgctg gagcgcgacc actggtactt cccctgccac tttgtgcgcg acgaggtgat ggccgggtcg ctggtcagcg acggctgctc gcagctcctc aaggtgtaca tgctgtggct cggcctgcac acgaccgtgg gcgcgttcga ctttcgtccc gtgagcgggc acgccaacaa ggtgcggtgc cgcgggcaga tctcaccgca caagggcaag ctcgtgtacg tgatggagat caaggaaatg ggctttgacg cgaagacggg cgatccgttt gcgatcgcgg acgtggacat catcgacgtc aacttcgagg agggacaggc gtttgcggga gtggaagacc tgcacagcta cggccagggc gacctccgca agaagatcgt cgtcgacttc aagggcatcg cgctctccct gcagaagcgg aaggagcagc agaaggaaag catgaccgtg < 210 > 38 < 211 > 450 < 212 > PRT <213> Schizochytrium species <400> 38

Met Thr Ser Ser Lys Lys Thr Pro Val Trp Glu Met Ser Lys Glu Glu 15 10 15Met Thr Ser Ser Lys Lys Thr Pro Val Trp Glu Met Ser Lys Glu Glu 15 10 15

Leu Leu Asp Gly Lys Thr Val Val Phe Asp Tyr Asn Glu Leu Leu Glu 20 25 30Leu Leu Asp Gly Lys Thr Val Val Phe Asp Tyr Asn Glu Leu Leu Glu 20 25 30

Phe Ala Glu Gly Asp Val Gly Gin Val Phe Gly Pro Glu Phe Asp lie 35 40 45 lie Asp Lys Tyr Arg Arg Arg Val Arg Leu Pro Ala Arg Glu Tyr Leu 50 55 60Phe Ala Glu Gly Asp Val Gly Gin Val Phe Gly Pro Glu Phe Asp lie 35 40 45 lie Asp Lys Tyr Arg Arg Arg Val Arg Leu Pro Ala Arg Glu Tyr Leu 50 55 60

Leu Val Ser Arg Val Thr Leu Met Asp Ala Glu Val Asn Asn Phe Arg 65 70 75 80Leu Val Ser Arg Val Thr Leu Met Asp Ala Glu Val Asn Asn Phe Arg 65 70 75 80

Val Gly Ser Arg Met Val Thr Glu Tyr Asp Val Pro Val Asn Gly Glu 85 90 95Val Gly Ser Arg Met Val Thr Glu Tyr Asp Val Pro Val Asn Gly Glu 85 90 95

Leu Ser Glu Gly Gly Asp Val Pro Trp Ala Val Leu Val Glu Ser Gly 100 105 110Leu Ser Glu Gly Gly Asp Val Pro Trp Ala Val Leu Val Glu Ser Gly 100 105 110

Gin Cys Asp Leu Met Leu lie Ser Tyr Met Gly lie Asp Phe Gin Cys 115 120 125Gin Cys Asp Leu Met Leu lie Ser Tyr Met Gly lie Asp Phe Gin Cys 115 120 125

Lys Gly Asp Arg Val Tyr Arg Leu Leu Asn Thr Ser Leu Thr Phe Phe 130 135 140Lys Gly Asp Arg Val Tyr Arg Leu Leu Asn Thr Ser Leu Thr Phe Phe 130 135 140

Gly Val Ala His Glu Gly Glu Thr Leu Val Tyr Asp lie Arg Val Thr 145 150 155 160 55 201038734Gly Val Ala His Glu Gly Glu Thr Leu Val Tyr Asp lie Arg Val Thr 145 150 155 160 55 201038734

Gly Phe Ala Lys Gly Ala Gly Gly Glu lie Ser Met Phe Phe Phe Glu 165 17〇 175Gly Phe Ala Lys Gly Ala Gly Gly Glu lie Ser Met Phe Phe Phe Glu 165 17〇 175

Tyr Asp Cys Phe Val Asp Gly Arg Leu Leu lie Glu Met Arg Asp Gly 180 185 190Tyr Asp Cys Phe Val Asp Gly Arg Leu Leu lie Glu Met Arg Asp Gly 180 185 190

Cys Ala Gly Phe Phe Thr Asp Ala Glu Leu Ala Ala Gly Lys Gly Val 195 200 205Cys Ala Gly Phe Phe Thr Asp Ala Glu Leu Ala Ala Gly Lys Gly Val 195 200 205

Leu Lys Thr Lys Ala Glu Leu Ala Ala Arg Ala Gin lie Gin Lys Gin 210 215 220Leu Lys Thr Lys Ala Glu Leu Ala Ala Arg Ala Gin lie Gin Lys Gin 210 215 220

Asp lie Ala Pro Phe Ala Pro Ala Pro Cys Ser His Lys Thr Ser Leu 225 230 235 240Asp lie Ala Pro Phe Ala Pro Ala Pro Cys Ser His Lys Thr Ser Leu 225 230 235 240

Asp Ala Arg Glu Met Arg Leu Leu Val Asp Arg Gin Trp Ala Arg Val 245 250 255Asp Ala Arg Glu Met Arg Leu Leu Val Asp Arg Gin Trp Ala Arg Val 245 250 255

Phe Gly Ser Gly Met Ala Gly lie Asp Tyr Lys Leu Cys Ala Arg Lys 260 265 270Phe Gly Ser Gly Met Ala Gly lie Asp Tyr Lys Leu Cys Ala Arg Lys 260 265 270

Met Leu Met 工le Asp Arg Val Thr His Leu Asp Pro Arg Gly Gly Ala 275 280 285Met Leu Met work le Asp Arg Val Thr His Leu Asp Pro Arg Gly Gly Ala 275 280 285

His Gly Leu Gly Leu Leu lie Gly Glu Lys Val Leu Glu Arg Asp His 290 295 300His Gly Leu Gly Leu Leu lie Gly Glu Lys Val Leu Glu Arg Asp His 290 295 300

Trp Tyr Phe Pro Cys His Phe Val Arg Asp Glu Val Met Ala Gly Ser 305 310 315 320Trp Tyr Phe Pro Cys His Phe Val Arg Asp Glu Val Met Ala Gly Ser 305 310 315 320

Leu Val Ser Asp Gly Cys Ser Gin Leu Leu Lys Val Tyr Met Leu Trp 325 330 335Leu Val Ser Asp Gly Cys Ser Gin Leu Leu Lys Val Tyr Met Leu Trp 325 330 335

Leu Gly Leu His Thr Thr Val Gly Ala Phe Asp Phe Arg Pro Val Ser 340 345 350Leu Gly Leu His Thr Thr Val Gly Ala Phe Asp Phe Arg Pro Val Ser 340 345 350

Gly His Ala Asn Lys Val Arg Cys Arg Gly Gin lie Ser Pro His Lys 355 360 365Gly His Ala Asn Lys Val Arg Cys Arg Gly Gin lie Ser Pro His Lys 355 360 365

Gly Lys Leu Val Tyr Val Met Glu lie Lys Glu Met Gly Phe Asp Ala 370 375 380Gly Lys Leu Val Tyr Val Met Glu lie Lys Glu Met Gly Phe Asp Ala 370 375 380

Lys Thr Gly Asp Pro Phe Ala lie Ala Asp Val Asp lie lie Asp Val 385 390 395 400Lys Thr Gly Asp Pro Phe Ala lie Ala Asp Val Asp lie lie Asp Val 385 390 395 400

Asn Phe Glu Glu Gly Gin Ala Phe Ala Gly Val Glu Asp Leu His Ser 405 410 415Asn Phe Glu Glu Gly Gin Ala Phe Ala Gly Val Glu Asp Leu His Ser 405 410 415

Tyr Gly Gin Gly Asp Leu Arg Lys Lys 工le Val Val Asp Phe Lys Gly 420 425 430 lie Ala Leu Ser Leu Gin Lys Arg Lys Glu Gin Gin Lys Glu Ser Met 435 440 445Tyr Gly Gin Gly Asp Leu Arg Lys Lys Le Val Val Asp Phe Lys Gly 420 425 430 lie Ala Leu Ser Leu Gin Lys Arg Lys Glu Gin Gin Lys Glu Ser Met 435 440 445

Thr Val 450 56 60 60Thr Val 450 56 60 60

Ο 201038734 <210> 39 <211> 1200 <212> DNA <213> 裂殖壺鏟物種 <400> 39 tacccgccgc gggcggtgtg cttctcgccg ttccccaaca acccgcttga caacgaccac acgccgggcc agatgccgtt gacctggttc aacatgtccg aattcatgtg cggcaaagtg tccaactgcc tgggccccga gtttgcgcgc ttcgacgcga gcaagacgag ccgcagcccg gcctttgacc tggcgctcgt gacgcgggtg acgagcgtgg cggacatgga gcacgggccg ttctacaacg tggacgtcaa cccgggccag ggcacgatgg tgggcgagtt cgactgtccc gcggacgcgt ggttcttcgg cgcctcgagc cgcgacgacc acatgccgta ctcgatcctg atggagatcg cgctgcagac gtcgggcgtc ctcacctcgg tgctcaaggc gccgctgacg atggacaagg acgacatcct cttccgcaac ctcgacgcag acgccgagct cgtgggcgac gccatgccgg acgtgcgcgg caagacgatc cgcaacttca ccaagtgcac aggctacagc atgctcggca agatgggcat ccaccgcttc acctttgagc tcagcgtcga cggcgccgtc ttctacaagg gcagcacctc gtttggctgg ttcgtccccg aggtcttcga gtcgcagacc ggtctcgaca acggcaagcc gcgcctgcct tggtaccgcg agaacaacgt cgccgtcgac acgctctccg cgcccgcctc cgcttcctcc gcgcaaggtc agctgcagct gcagcgacgc gggtcgcagg cgcagttcct ggacacaatc cacctggcgg gcagcggcgc cggcgtgcac ggccagggct acgcgcacgg ggagaaggcc gtgaacaagc aagattggtt cttctcgtgc cacttctggt tcgaccccgt gatgcccggg tccctgggca tcgagtcgat gttccagctc gtcgaggcgt ggtgcgtgaa gcagggactc gcggcgcggc acggcatcgc tcacccagtg ttcgcgcacg cgcccggggc cacgagctgg aagtaccgcg ggcagctaac ccccaagaac gaccgcatgg acagcgaggt gcacatcaag tcggtggcgg ccttctcctc ctgggtcgac gtcgtcgcgg acgggttcct cttcgtcgac ggcctccgcg tctactcggc agacaacctc <210> 40 <211〉 400 <212> PRT <213〉 裂殖壺菌物種 <400> 40Ο 201038734 < 210 > 39 < 211 > 1200 < 212 > DNA < 213 > Schizochytrium shovel species < 400 > 39 tacccgccgc gggcggtgtg cttctcgccg ttccccaaca acccgcttga caacgaccac acgccgggcc agatgccgtt gacctggttc aacatgtccg aattcatgtg cggcaaagtg tccaactgcc tgggccccga gtttgcgcgc ttcgacgcga gcaagacgag ccgcagcccg gcctttgacc tggcgctcgt gacgcgggtg acgagcgtgg cggacatgga gcacgggccg ttctacaacg tggacgtcaa cccgggccag ggcacgatgg tgggcgagtt cgactgtccc gcggacgcgt ggttcttcgg cgcctcgagc cgcgacgacc acatgccgta ctcgatcctg atggagatcg cgctgcagac gtcgggcgtc ctcacctcgg tgctcaaggc gccgctgacg atggacaagg acgacatcct cttccgcaac ctcgacgcag acgccgagct cgtgggcgac gccatgccgg acgtgcgcgg caagacgatc cgcaacttca ccaagtgcac aggctacagc atgctcggca agatgggcat ccaccgcttc acctttgagc tcagcgtcga cggcgccgtc ttctacaagg gcagcacctc gtttggctgg ttcgtccccg aggtcttcga gtcgcagacc ggtctcgaca acggcaagcc gcgcctgcct Tggtaccgcg agaacaacgt cgccgtcgac acgctctccg cgcccgcctc cgcttcctcc gcgcaaggtc agctgcagct gcagcgacgc gggtcgcagg cgcagttcct ggacacaatc ca cctggcgg gcagcggcgc cggcgtgcac ggccagggct acgcgcacgg ggagaaggcc gtgaacaagc aagattggtt cttctcgtgc cacttctggt tcgaccccgt gatgcccggg tccctgggca tcgagtcgat gttccagctc gtcgaggcgt ggtgcgtgaa gcagggactc gcggcgcggc acggcatcgc tcacccagtg ttcgcgcacg cgcccggggc cacgagctgg aagtaccgcg ggcagctaac ccccaagaac gaccgcatgg acagcgaggt gcacatcaag tcggtggcgg ccttctcctc ctgggtcgac gtcgtcgcgg acgggttcct cttcgtcgac ggcctccgcg tctactcggc agacaacctc < 210 > 40 < 211> 400 <212> PRT <213> Schizochytrium species <400> 40

Tyr Pro Pro Arg Ala Val Cys Phe Ser Pro Phe Pro Asn Asn Pro Leu 15 10 15Tyr Pro Pro Arg Ala Val Cys Phe Ser Pro Phe Pro Asn Asn Pro Leu 15 10 15

Asp Asn Asp His Thr Pro Gly Gin Met Pro Leu Thr Trp Phe Asn Met 20 25 30Asp Asn Asp His Thr Pro Gly Gin Met Pro Leu Thr Trp Phe Asn Met 20 25 30

Ser Glu Phe Met Cys Gly Lys Val Ser Asn Cys Leu Gly Pro Glu Phe 35 40 45Ser Glu Phe Met Cys Gly Lys Val Ser Asn Cys Leu Gly Pro Glu Phe 35 40 45

Ala Arg Phe Asp Ala Ser Lys Thr Ser Arg Ser Pro Ala Phe Asp Leu 50 55 60Ala Arg Phe Asp Ala Ser Lys Thr Ser Arg Ser Pro Ala Phe Asp Leu 50 55 60

Ala Leu Val Thr Arg Val Thr Ser Val Ala Asp Met Glu His Gly Pro 65 70 75 80 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 57 201038734Ala Leu Val Thr Arg Val Thr Ser Val Ala Asp Met Glu His Gly Pro 65 70 75 80 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 57 201038734

Phe Tyr Asn Val Asp Val Asn Pro Gly Gin Gly Thr Met Val Gly Glu 85 90 95Phe Tyr Asn Val Asp Val Asn Pro Gly Gin Gly Thr Met Val Gly Glu 85 90 95

Phe Asp Cys Pro Ala Asp Ala Trp Phe Phe Gly Ala Ser Ser Arg Asp 100 105 110Phe Asp Cys Pro Ala Asp Ala Trp Phe Phe Gly Ala Ser Ser Arg Asp 100 105 110

Asp His Met Pro Tyr Ser lie Leu Met Glu lie Ala Leu Gin Thr Ser 115 120 125Asp His Met Pro Tyr Ser lie Leu Met Glu lie Ala Leu Gin Thr Ser 115 120 125

Gly Val Leu Thr Ser Val Leu Lys Ala Pro Leu Thr Met Asp Lys Asp 130 135 140Gly Val Leu Thr Ser Val Leu Lys Ala Pro Leu Thr Met Asp Lys Asp 130 135 140

Asp lie Leu Phe Arg Asn Leu Asp Ala Asp Ala Glu Leu Val Gly Asp 145 150 155 160Asp lie Leu Phe Arg Asn Leu Asp Ala Asp Ala Glu Leu Val Gly Asp 145 150 155 160

Ala Met Pro Asp Val Arg Gly Lys Thr lie Arg Asn Phe Thr Lys Cys 165 170 175Ala Met Pro Asp Val Arg Gly Lys Thr lie Arg Asn Phe Thr Lys Cys 165 170 175

Thr Gly Tyr Ser Met Leu Gly Lys Met Gly lie His Arg Phe Thr Phe 180 185 190Thr Gly Tyr Ser Met Leu Gly Lys Met Gly lie His Arg Phe Thr Phe 180 185 190

Glu Leu Ser Val Asp Gly Ala Val Phe Tyr Lys Gly Ser Thr Ser Phe 195 200 205Glu Leu Ser Val Asp Gly Ala Val Phe Tyr Lys Gly Ser Thr Ser Phe 195 200 205

Gly Trp Phe Val Pro Glu Val Phe Glu Ser Gin Thr Gly Leu Asp Asn 210 215 220Gly Trp Phe Val Pro Glu Val Phe Glu Ser Gin Thr Gly Leu Asp Asn 210 215 220

Gly Lys Pro Arg Leu Pro Trp Tyr Arg Glu Asn Asn Val Ala Val Asp 225 230 235 240Gly Lys Pro Arg Leu Pro Trp Tyr Arg Glu Asn Asn Val Ala Val Asp 225 230 235 240

Thr Leu Ser Ala Pro Ala Ser Ala Ser Ser Ala Gin Gly Gin Leu Gin 245 250 255Thr Leu Ser Ala Pro Ala Ser Ala Ser Ser Ala Gin Gly Gin Leu Gin 245 250 255

Leu Gin Arg Arg Gly Ser Gin Ala Gin Phe Leu Asp Thr lie His Leu 260 265 270Leu Gin Arg Arg Gly Ser Gin Ala Gin Phe Leu Asp Thr lie His Leu 260 265 270

Ala Gly Ser Gly Ala Gly Val His Gly Gin Gly Tyr Ala His Gly Glu 275 280 285Ala Gly Ser Gly Ala Gly Val His Gly Gin Gly Tyr Ala His Gly Glu 275 280 285

Lys Ala Val Asn Lys Gin Asp Trp Phe Phe Ser Cys His Phe Trp Phe 290 295 300Lys Ala Val Asn Lys Gin Asp Trp Phe Phe Ser Cys His Phe Trp Phe 290 295 300

Asp Pro Val Met Pro Gly Ser Leu Gly lie Glu Ser Met Phe Gin Leu 305 310 315 320Asp Pro Val Met Pro Gly Ser Leu Gly lie Glu Ser Met Phe Gin Leu 305 310 315 320

Val Glu Ala Trp Cys Val Lys Gin Gly Leu Ala Ala Arg His Gly lie 325 330 335Val Glu Ala Trp Cys Val Lys Gin Gly Leu Ala Ala Arg His Gly lie 325 330 335

Ala His Pro Val Phe Ala His Ala Pro Gly Ala Thr Ser Trp Lys Tyr 340 345 350Ala His Pro Val Phe Ala His Ala Pro Gly Ala Thr Ser Trp Lys Tyr 340 345 350

Arg Gly Gin Leu Thr Pro Lys Asn Asp Arg Met Asp Ser Glu Val His 355 360 365 工le Lys Ser Val Ala Ala Phe Ser Ser Trp Val Asp Val Val Ala Asp 370 375 , 380 58 201038734Arg Gly Gin Leu Thr Pro Lys Asn Asp Arg Met Asp Ser Glu Val His 355 360 365 work le Lys Ser Val Ala Ala Phe Ser Ser Trp Val Asp Val Val Ala Asp 370 375 , 380 58 201038734

Gly Phe Leu Phe Val Asp Gly Leu Arg Val Tyr Ser Ala Asp Asn Leu 385 390 395 400Gly Phe Leu Phe Val Asp Gly Leu Arg Val Tyr Ser Ala Asp Asn Leu 385 390 395 400

<210> 41 <211> 1353 <212> DNA <213> 裂殖壺菌物種 <400> 41 cagctggacg cggggagcga ggtgcccgcc tgcgcggtga gcgacctggg cgataggggc 60 ttcatggaga cgtacggggt ggtggcgccg ctgtacagcg gggcgatggc caagggcatc 120 gcgtcggcgg acctggtgat cgcgatgggc cagcgcaaga tgctggggtc gtttggcgcg 180 ggcgggctcc cgatgcacgt cgtgcgcgcg gggattgaga agatccaggc agcgctgcca 240 gcggggccat acgcggtcaa cctgattcac tcgccttttg acgccaacct ggagaagggc 300 aacgtggacc tcttcctgga gaagggcgtg cgcgtcgtgg aggcgtcggc cttcatggag 360 ctcacgcccc aggtggtgcg ctaccgcgcg acgggcctct ctcgcgacgc gcgcggcggc 420 tccgtgcgca cggcccacaa gatcatcggc aaggtcagcc gcaccgagct ggccgagatg 480 tttatccggc ccgcgccgca agccattctc gacaagcttg tggcgtccgg cgagatcacc 540 cccgagcagg cggcgctggc gctcgaggtg cccatggcgg acgacatcgc cgtcgaggcc 600 gattcgggcg ggcacaccga caaccgcccc atccacgtca tcctgcccct catcctcagc 660 ctgcgcaacc gcctccagcg cgagctcaag taccctgcgc gacaccgcgt gcgcgtcggc 720 gccgggggcg gcatcgggtg cccgcaagcg gctctgggcg ccttccacat gggcgccgcg 780 tttgtggtga cgggcacggt caaccagctg agccggcagg ccgggacatg cgacaatgtg 840 cggcggcagc tgtcgcgcgc gacgtactcg gacatcacga tggcgccggc ggcggacatg 900 ttcgagcagg gcgtcgagct gcaggtgctc aagaagggca cgatgtttcc ctcgcgcgcc 960 aagaagctgt tcgagctgtt tcacaagtac gactcgttcg aggcgatgcc ggcggacgag 1020 ctggcgcgcg tcgagaagcg catcttcagc aagtcactcg ccgaggtgtg ggccgagacc 1080 aaggacttct acatcacgcg gctcaacaac ccggagaaga tccgcaaggc ggagaacgag 1140 gaccccaagc tcaagatgtc actctgcttc cgctggtacc tcgggctcag ctcgttctgg 1200 gccaacaacg gcatcgcgga ccgcacgatg gactaccaga tctggtgcgg ccctgccatc 1260 ggcgccttca acgacttcat cgccgactcg tacctcgacg tggccgtctc gggcgagttc 1320 cccgacgtcg tgcagatcaa cctgcagatc ctg 1353 <210> 42 <211> 451 <212> PRT <213〉 裂殖壺菌物種 <400> 42≪ 210 > 41 < 211 > 1353 < 212 > DNA < 213 > Schizochytrium species < 400 > 41 cagctggacg cggggagcga ggtgcccgcc tgcgcggtga gcgacctggg cgataggggc 60 ttcatggaga cgtacggggt ggtggcgccg ctgtacagcg gggcgatggc caagggcatc 120 gcgtcggcgg acctggtgat cgcgatgggc cagcgcaaga tgctggggtc gtttggcgcg 180 ggcgggctcc cgatgcacgt cgtgcgcgcg gggattgaga agatccaggc agcgctgcca 240 gcggggccat acgcggtcaa cctgattcac tcgccttttg acgccaacct ggagaagggc 300 aacgtggacc tcttcctgga gaagggcgtg cgcgtcgtgg aggcgtcggc cttcatggag 360 ctcacgcccc aggtggtgcg ctaccgcgcg acgggcctct ctcgcgacgc gcgcggcggc 420 tccgtgcgca cggcccacaa gatcatcggc aaggtcagcc gcaccgagct ggccgagatg 480 tttatccggc ccgcgccgca agccattctc gacaagcttg tggcgtccgg cgagatcacc 540 cccgagcagg cggcgctggc gctcgaggtg cccatggcgg acgacatcgc cgtcgaggcc 600 gattcgggcg Ggcacaccga caaccgcccc atccacgtca tcctgcccct catcctcagc 660 ctgcgcaacc gcctccagcg cgagctcaag taccctgcgc gacaccgcgt gcgcgtcggc 720 gccggggggg gcatcgggtg cccgcaagcg gctctgggcg ccttc cacat gggcgccgcg 780 tttgtggtga cgggcacggt caaccagctg agccggcagg ccgggacatg cgacaatgtg 840 cggcggcagc tgtcgcgcgc gacgtactcg gacatcacga tggcgccggc ggcggacatg 900 ttcgagcagg gcgtcgagct gcaggtgctc aagaagggca cgatgtttcc ctcgcgcgcc 960 aagaagctgt tcgagctgtt tcacaagtac gactcgttcg aggcgatgcc ggcggacgag 1020 ctggcgcgcg tcgagaagcg catcttcagc aagtcactcg ccgaggtgtg ggccgagacc 1080 aaggacttct acatcacgcg gctcaacaac ccggagaaga tccgcaaggc ggagaacgag 1140 gaccccaagc tcaagatgtc actctgcttc cgctggtacc tcgggctcag ctcgttctgg 1200 gccaacaacg gcatcgcgga ccgcacgatg gactaccaga tctggtgcgg ccctgccatc 1260 ggcgccttca acgacttcat cgccgactcg tacctcgacg tggccgtctc gggcgagttc 1320 cccgacgtcg tgcagatcaa cctgcagatc ctg 1353 < 210 > 42 < 211 > 451 < 212 > PRT < 213> Schizochytrium species < 400 > 42

Gin Leu Asp Ala Gly Ser Glu Val Pro Ala Cys Ala Val Ser Asp Leu 15 10 15Gin Leu Asp Ala Gly Ser Glu Val Pro Ala Cys Ala Val Ser Asp Leu 15 10 15

Gly Asp Arg Gly Phe Met 20Gly Asp Arg Gly Phe Met 20

Ser Gly Ala Met Ala Lys 35Ser Gly Ala Met Ala Lys 35

Glu Thr Tyr Gly Val Val Ala Pro Leu Tyr 25 30Glu Thr Tyr Gly Val Val Ala Pro Leu Tyr 25 30

Gly lie Ala Ser Ala Asp Leu Val lie Ala 40 45 59 201038734Gly lie Ala Ser Ala Asp Leu Val lie Ala 40 45 59 201038734

Met Gly Gin Arg Lys Met Leu Gly Ser Phe Gly Ala Gly Gly Leu Pro 50 55 60Met Gly Gin Arg Lys Met Leu Gly Ser Phe Gly Ala Gly Gly Leu Pro 50 55 60

Met His Val Val Arg Ala Gly lie Glu Lys lie Gin Ala Ala Leu Pro 65 70 75 80Met His Val Val Arg Ala Gly lie Glu Lys lie Gin Ala Ala Leu Pro 65 70 75 80

Ala Gly Pro Tyr Ala Val Asn Leu lie His Ser Pro Phe Asp Ala Asn 85 90 95Ala Gly Pro Tyr Ala Val Asn Leu lie His Ser Pro Phe Asp Ala Asn 85 90 95

Leu Glu Lys Gly Asn Val Asp Leu Phe Leu Glu Lys Gly Val Arg Val 100 105 110Leu Glu Lys Gly Asn Val Asp Leu Phe Leu Glu Lys Gly Val Arg Val 100 105 110

Val Glu Ala Ser Ala Phe Met Glu Leu Thr Pro Gin Val Val Arg Tyr 115 120 125Val Glu Ala Ser Ala Phe Met Glu Leu Thr Pro Gin Val Val Arg Tyr 115 120 125

Arg Ala Thr Gly Leu Ser Arg Asp Ala Arg Gly Gly Ser Val Arg Thr 130 135 140 e _—l I s Ly s •1 H a 5 1 4 Δ -~_ lie Gly Lys Val Ser Arg Thr Glu Leu Ala Glu Met 150 155 160Arg Ala Thr Gly Leu Ser Arg Asp Ala Arg Gly Gly Ser Val Arg Thr 130 135 140 e _-l I s Ly s •1 H a 5 1 4 Δ -~_ lie Gly Lys Val Ser Arg Thr Glu Leu Ala Glu Met 150 155 160

Phe 工le Arg Pro Ala Pro Gin Ala lie Leu Asp Lys Leu Val Ala Ser 165 170 175Phe work le Arg Pro Ala Pro Gin Ala lie Leu Asp Lys Leu Val Ala Ser 165 170 175

Gly Glu lie Thr Pro Glu Gin Ala Ala Leu Ala Leu Glu Val Pro Met 180 185 190Gly Glu lie Thr Pro Glu Gin Ala Ala Leu Ala Leu Glu Val Pro Met 180 185 190

Ala Asp Asp lie Ala Val Glu Ala Asp Ser Gly Gly His Thr Asp Asn 195 200 205Ala Asp Asp lie Ala Val Glu Ala Asp Ser Gly Gly His Thr Asp Asn 195 200 205

g r A oo r.—_ p 2 a V s i H eg r A oo r.—_ p 2 a V s i H e

u 5 el L2 e _—_ I r pu 5 el L2 e _—_ I r p

u e Lu e L

r o Θ 2 s 2 u e Lr o Θ 2 s 2 u e L

Leu Arg Asn ArgLeu Arg Asn Arg

Leu Gin Arg Glu Leu Lys Tyr Pro Ala Arg His Arg Val Arg Val Gly 225 230 235 240Leu Gin Arg Glu Leu Lys Tyr Pro Ala Arg His Arg Val Arg Val Gly 225 230 235 240

Ala Gly Gly Gly lie Gly Cys 245 A a o 15 A 2 n _—I G 〇 r pAla Gly Gly Gly lie Gly Cys 245 A a o 15 A 2 n _—I G 〇 r p

s i H e 5 h 5 p 2 a _—I A V G u e Ls i H e 5 h 5 p 2 a _—I A V G u e L

Met Gly Ala Ala Phe Val Val Thr Gly Thr Val Asn Gin Leu Ser Arg 260 265 270Met Gly Ala Ala Phe Val Val Thr Gly Thr Val Asn Gin Leu Ser Arg 260 265 270

Gin Ala Gly Thr Cys Asp Asn Val Arg Arg Gin Leu Ser Arg Ala Thr 275 280 285Gin Ala Gly Thr Cys Asp Asn Val Arg Arg Gin Leu Ser Arg Ala Thr 275 280 285

Tyr Ser Asp lie Thr Met Ala Pro Ala Ala Asp Met Phe Glu Gin Gly 290 295 300Tyr Ser Asp lie Thr Met Ala Pro Ala Ala Asp Met Phe Glu Gin Gly 290 295 300

Val Glu Leu Gin Val Leu Lys Lys Gly Thr Met Phe Pro Ser Arg Ala 305 310 315 320Val Glu Leu Gin Val Leu Lys Lys Gly Thr Met Phe Pro Ser Arg Ala 305 310 315 320

Lys Lys Leu Phe Glu Leu Phe His Lys Tyr Asp Ser Phe Glu Ala Met 325 330 335Lys Lys Leu Phe Glu Leu Phe His Lys Tyr Asp Ser Phe Glu Ala Met 325 330 335

Pro Ala Asp Glu Leu Ala Arg Val Glu Lys Arg lie Phe Ser Lys Ser 60 201038734 340 345 350Pro Ala Asp Glu Leu Ala Arg Val Glu Lys Arg lie Phe Ser Lys Ser 60 201038734 340 345 350

Leu Ala Glu Val Trp Ala Glu Thr Lys Asp Phe Tyr lie Thr Arg Leu 355 360 365Leu Ala Glu Val Trp Ala Glu Thr Lys Asp Phe Tyr lie Thr Arg Leu 355 360 365

Asn Asn Pro Glu Lys lie Arg Lys Ala Glu Asn Glu Asp Pro Lys Leu 370 375 380 s V c u e L r e s t e M s 5 y8 L 3Asn Asn Pro Glu Lys lie Arg Lys Ala Glu Asn Glu Asp Pro Lys Leu 370 375 380 s V c u e L r e s t e M s 5 y8 L 3

Phe Arg Trp Tyr Leu Gly Leu Ser Ser Phe Trp 390 395 400Phe Arg Trp Tyr Leu Gly Leu Ser Ser Phe Trp 390 395 400

Ala Asn Asn Gly lie Ala Asp Arg Thr Met Asp Tyr Gin lie Trp Cys 405 410 415Ala Asn Asn Gly lie Ala Asp Arg Thr Met Asp Tyr Gin lie Trp Cys 405 410 415

Gly Pro Ala lie Gly Ala Phe Asn Asp Phe lie Ala Asp Ser Tyr Leu 420 425 430 oGly Pro Ala lie Gly Ala Phe Asn Asp Phe lie Ala Asp Ser Tyr Leu 420 425 430 o

Asp Val Ala Val Ser Gly Glu Phe Pro Asp Val Val Gin lie Asn Leu 435 440 445Asp Val Ala Val Ser Gly Glu Phe Pro Asp Val Val Gin lie Asn Leu 435 440 445

Gin lie Leu 450 <210> 43 <211> 4 <212> PRT <213> 裂殖壺菌物種 <220> <221> 其他特徵 <222> (2)..(2) <223> Xaa可為任何 <400> 43 Asp Xaa Ala Cys 1 <210〉 44 <211> 4 <212> PRT <213〉 裂殖壺菌物種 <400> 44 Gly Phe Gly Gly 1 <210> 45 <211> 12 <212> DNA <213> 裂殖壺菌物種 <400> 45 gggtttggcg gt <21〇> 46 <211> 5 <212> PRT <213> 裂殖壺菌物種 <4〇0> 46 61 12 201038734Gin lie Leu 450 <210> 43 <211> 4 <212> PRT <213> Schizochytrium species <220><221> Other characteristics <222> (2)..(2) <223> Xaa can be any <400> 43 Asp Xaa Ala Cys 1 <210> 44 <211> 4 <212> PRT <213> Schizochytrium species <400> 44 Gly Phe Gly Gly 1 <210> 45 <211> 12 <212> DNA <213> Schizochytrium species <400> 45 gggtttggcg gt <21〇> 46 <211> 5 <212> PRT <213> Schizochytrium species <4〇0> 46 61 12 201038734

Gly Phe Ser Leu Gly 1 5 <210> 47 <211> 5 <212> PRT <213> 裂殖壺菌物種 <400> 47Gly Phe Ser Leu Gly 1 5 <210> 47 <211> 5 <212> PRT <213> Schizochytrium species <400>

Leu Gly lie Asp Ser 1 5 <210> 48 <211> 903 <212> DNA <213〉 裂?i壺菌物種 <400> 48 ctgtcagacc ccgacgtggc cgtgcgcgag tctggttact ccgcctcggg ccagcgctgc 60 acgacaacta cgaagtcgct gactacgggc aagccgcacc agccgatctc ctcgtcggac 120 ctctttctgg tgtcgggcgg cgcgcgcggc atcaccccgc tgtgcgtgcg cgagctggcg 180 cagcgcgtgg gcggcggcac gtacgtgctc atcggccgct cggagctgcc cacgacggag 240 cctgcctggg cggtcggcgt ggagtctggc aagccgctgg agaaggccgc gctggcgttc 300 ctgaaggcgg agtttgcagc gggccgcggg gccaagccga cgccgatgct gcacaagaag 360 ctcgtgggcg ccgtggtcgg agcgcgcgag gtgcgagcct cgctcgccga gatcactgca 420 cagggcgcca cggctgtgta cgagtcgtgc gacgtgagct ctgccgccaa ggtgcgtgag 480 atggtagagc gcgtgcagca gcagggcggg cggcgcgtgt cgggcgtgtt ccacgcgtcg 540 ggcgtgctgc gcgacaagct cgtggagaac aagtcgctgg cggacttcag cgccgtgtac 600 gacaccaagg tgggcggcct catcaacctg ctggcctgcg tggacctggc gcagctgcgt 660 cacctcgtgc tcttcagctc gctcgcgggc ttccacggca acgtcgggca gtcggactac 720 gcaatggcca acgaggcgct caacaagctg gcggcgcacc tgtcggcggt gcacccgcag 7 80 ctgtgcgcgc gctcgatctg cttcggaccg tgggacggcg gcatggtgac ccccgcgctc 840 aaggccaact tcatccgcat gggcatccag atcatcccgc gccaaggcgg cgcgcagacc 900 gtc 903 <210> 49 <211> 301 <212> PRT <213> 裂殖壺菌物種 <400> 49Leu Gly lie Asp Ser 1 5 <210> 48 <211> 903 <212> DNA <213> cleavage? I chymogen species <400> 48 ctgtcagacc ccgacgtggc cgtgcgcgag tctggttact ccgcctcggg ccagcgctgc 60 acgacaacta cgaagtcgct gactacgggc aagccgcacc agccgatctc ctcgtcggac 120 ctctttctgg tgtcgggcgg cgcgcgcggc atcaccccgc tgtgcgtgcg cgagctggcg 180 cagcgcgtgg gcggcggcac gtacgtgctc atcggccgct cggagctgcc cacgacggag 240 cctgcctggg cggtcggcgt ggagtctggc aagccgctgg agaaggccgc gctggcgttc 300 ctgaaggcgg agtttgcagc gggccgcggg gccaagccga cgccgatgct gcacaagaag 360 ctcgtgggcg ccgtggtcgg agcgcgcgag gtgcgagcct cgctcgccga gatcactgca 420 cagggcgcca cggctgtgta cgagtcgtgc gacgtgagct ctgccgccaa ggtgcgtgag 480 atggtagagc gcgtgcagca gcagggcggg cggcgcgtgt cgggcgtgtt ccacgcgtcg 540 Ggcgtgctgc gcgacaagct cgtggagaac aagtcgctgg cggacttcag cgccgtgtac 600 gacaccaagg tgggcggcct catcaacctg ctggcctgcg tggacctggc gcagctgcgt 660 cacctcgtgc tcttcagctc gctcgcgggc ttccacggca acgtcgggca gtcggactac 720 gcaatggcca acgaggcgct caacaagctg gcggcgcacc tgtcggcggt gcacccgcag 7 80 ctgtgcgcgc gctcgatctg cttcggaccg tgggacggcg gcatggtgac ccccgcgctc 840 aaggccaact tcatccgcat gggcatccag atcatcccgc gccaaggcgg cgcgcagacc 900 gtc 903 < 210 > 49 < 211 > 301 < 212 > PRT < 213 > Schizochytrium species < 400 > 49

Leu Ser Asp Pro Asp Val Ala Val Arg Glu Ser Gly Tyr Ser Ala Ser 1 5 10 15Leu Ser Asp Pro Asp Val Ala Val Arg Glu Ser Gly Tyr Ser Ala Ser 1 5 10 15

Gly Gin Arg Cys Thr Thr Thr Thr Lys Ser Leu Thr Thr Gly Lys Pro 20 25 30Gly Gin Arg Cys Thr Thr Thr Thr Lys Ser Leu Thr Thr Gly Lys Pro 20 25 30

His Gin Pro lie Ser Ser Ser Asp Leu Phe Leu Val Ser Gly Gly Ala 35 40 45His Gin Pro lie Ser Ser Ser Asp Leu Phe Leu Val Ser Gly Gly Ala 35 40 45

Arg Gly lie Thr Pro Leu Cys Val Arg Glu Leu Ala Gin Arg Val Gly 50 55 60 62 201038734Arg Gly lie Thr Pro Leu Cys Val Arg Glu Leu Ala Gin Arg Val Gly 50 55 60 62 201038734

Gly Gly Thr Tyr Val Leu lie Gly Arg Ser Glu Leu Pro Thr Thr Glu 65 70 75 80Gly Gly Thr Tyr Val Leu lie Gly Arg Ser Glu Leu Pro Thr Thr Glu 65 70 75 80

Pro Ala Trp Ala Val Gly Val Glu Ser Gly Lys Pro Leu Glu Lys Ala 85 90 95Pro Ala Trp Ala Val Gly Val Glu Ser Gly Lys Pro Leu Glu Lys Ala 85 90 95

Ala Leu Ala Phe Leu Lys Ala Glu Phe Ala Ala Gly Arg Gly Ala Lys 100 105 110Ala Leu Ala Phe Leu Lys Ala Glu Phe Ala Ala Gly Arg Gly Ala Lys 100 105 110

Pro Thr Pro Met Leu His Lys Lys Leu Val Gly Ala Val Val Gly Ala 115 120 125Pro Thr Pro Met Leu His Lys Lys Leu Val Gly Ala Val Val Gly Ala 115 120 125

Arg Glu Val Arg Ala Ser Leu Ala Glu lie Thr Ala Gin Gly Ala Thr 130 135 140Arg Glu Val Arg Ala Ser Leu Ala Glu lie Thr Ala Gin Gly Ala Thr 130 135 140

Ala Val Tyr Glu Ser Cys Asp Val Ser Ser Ala Ala Lys Val Arg GluAla Val Tyr Glu Ser Cys Asp Val Ser Ser Ala Ala Lys Val Arg Glu

145 150 155 160145 150 155 160

Met Val Glu Arg Val Gin Gin Gin Gly Gly Arg Arg Val Ser Gly Val 165 170 175Met Val Glu Arg Val Gin Gin Gin Gly Gly Arg Arg Val Ser Gly Val 165 170 175

Phe His Ala Ser Gly Val Leu Arg Asp Lys Leu Val Glu Asn Lys Ser 180 185 190Phe His Ala Ser Gly Val Leu Arg Asp Lys Leu Val Glu Asn Lys Ser 180 185 190

Leu Ala Asp Phe Ser Ala Val Tyr Asp Thr Lys Val Gly Gly Leu lie 195 200 205Leu Ala Asp Phe Ser Ala Val Tyr Asp Thr Lys Val Gly Gly Leu lie 195 200 205

Asn Leu Leu Ala Cys Val Asp Leu Ala Gin Leu Arg His Leu Val Leu 210 215 220Asn Leu Leu Ala Cys Val Asp Leu Ala Gin Leu Arg His Leu Val Leu 210 215 220

Phe Ser Ser Leu Ala Gly Phe His Gly Asn Val Gly Gin Ser Asp Tyr 225 230 235 240Phe Ser Ser Leu Ala Gly Phe His Gly Asn Val Gly Gin Ser Asp Tyr 225 230 235 240

Ala Met Ala Asn Glu Ala Leu Asn Lys Leu Ala Ala His Leu Ser Ala 245 250 255Ala Met Ala Asn Glu Ala Leu Asn Lys Leu Ala Ala His Leu Ser Ala 245 250 255

Val His Pro Gin Leu Cys Ala Arg Ser He Cys Phe Gly Pro Trp AspVal His Pro Gin Leu Cys Ala Arg Ser He Cys Phe Gly Pro Trp Asp

Gly Gly Met Val Thr Pro Ala Leu Lys Ala Asn Phe lie Arg Met Gly 275 280 285Gly Gly Met Val Thr Pro Ala Leu Lys Ala Asn Phe lie Arg Met Gly 275 280 285

Ile Ile Ile Pro Arg Gln G1y G1y Ala Gln Thr Val 290 295 300 <210> 50 <211> 13 <212> PRT <213> 裂殖壺菌物種 <220> <221> 其他特徵 <222> (2)..(3) <223> Xaa可為任何天然存在的胺基酸 <220〉 <221> 其他特徵 63 201038734 <222〉 （5)..(7) <223> Xaa可為任何天然存在的胺基酸 > > > > 0 12 3 2 2 2 2 2 2 2 2 < < < < 其他特徵 (9)..(12) ^ Xaa寸為任何天然存在的胺基酸 <400> 50 Leu Xaa Xaa His Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Pro 15 10 <210> 51 <211〉 12 <212> DNA <213> 裂殖壺菌物種 <400> 51 ggctttggtg ga 12 <210> 52 <211> 5 <212> PRT <213〉 裂殖壺菌物種 > > > > 0 12 3 2 2 2 2 2 2 2 2 < < < < 其他特徵(2) . . (2) Xaa可為任何天然存在的胺基酸 <220> <221> 其他特徵 <222> (4) .(4) <223> Xaa _计為任何天然存在的胺基酸 <400> 52 Gly Xaa Ser Xaa Gly 1 5 <210> 53 <211> 5 <212> PRT <213> 裂殖壺菌物種 > > > > 0 12 3 2 2 2 2 2 2 2 2 < < < < 其他特徵 (2) ·· (3) Xaa可為任何天然存在的胺基酸 <400> 53Ile Ile Ile Pro Arg Gln G1y G1y Ala Gln Thr Val 290 295 300 <210> 50 <211> 13 <212> PRT <213> Schizochytrium species <220><221> Other characteristics <;222> (2)..(3) <223> Xaa can be any naturally occurring amino acid <220> <221> Other features 63 201038734 <222> (5)..(7) <;223> Xaa can be any naturally occurring amino acid >>>> 0 12 3 2 2 2 2 2 2 2 2 <<<< Other characteristics (9).. (12) ^ Xaa is any naturally occurring amino acid <400> 50 Leu Xaa Xaa His Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Pro 15 10 <210> 51 <211> 12 <212> DNA <213> Schizochytrium species <400> 51 ggctttggtg ga 12 <210> 52 <211> 5 <212> PRT <213> Schizochytrium species >>>> 0 12 3 2 2 2 2 2 2 2 2 <<<< Other characteristics (2) . (2) Xaa may be any naturally occurring amino acid <220><221> Other characteristics <222> (4) <223> Xaa _ is calculated as any naturally occurring amino acid <400 > 52 Gly Xaa Ser Xaa Gly 1 5 <210> 53 <211> 5 <212> PRT <213> Schizochytrium species>>>> 0 12 3 2 2 2 2 2 2 2 2 <<<< Other Features (2) · (3) Xaa can be any naturally occurring amino acid <400>

Phe Xaa Xaa His Phe 1 5 <210〉 <211> <212> <213> 54 2910 PRT 裂殖壺菌物種 <400> 54 Met Ala 1 Ala Arg Leu 5 lie Ala lie lie 20 Gly 25 10 15 30 64 201038734Phe Xaa Xaa His Phe 1 5 <210> <211><212><213> 54 2910 PRT Schizochytrium species <400> 54 Met Ala 1 Ala Arg Leu 5 lie Ala lie lie 20 Gly 25 10 15 30 64 201038734

Arg Glu Ser Trp Glu Thr lie Arg Ala Gly lie Asp Cys Leu Ser Asp 35 40 45Arg Glu Ser Trp Glu Thr lie Arg Ala Gly lie Asp Cys Leu Ser Asp 35 40 45

Leu Pro Glu Asp Arg Val Asp Val Thr Ala Tyr Phe Asp Pro Val Lys 50 55 60Leu Pro Glu Asp Arg Val Asp Val Thr Ala Tyr Phe Asp Pro Val Lys 50 55 60

Thr Thr Lys Asp Lys lie Tyr Cys Lys Arg Gly Gly Phe lie Pro Glu 65 70 75 80Thr Thr Lys Asp Lys lie Tyr Cys Lys Arg Gly Gly Phe lie Pro Glu 65 70 75 80

Tyr Asp Phe Asp Ala Arg Glu Phe Gly Leu Asn Met Phe Gin Met Glu 85 90 95Tyr Asp Phe Asp Ala Arg Glu Phe Gly Leu Asn Met Phe Gin Met Glu 85 90 95

Asp Ser Asp Ala Asn Gin Thr lie Ser Leu Leu Lys Val Lys Glu Ala 100 105 110Asp Ser Asp Ala Asn Gin Thr lie Ser Leu Leu Lys Val Lys Glu Ala 100 105 110

Leu Gin Asp Ala Gly lie Asp Ala Leu Gly Lys Glu Lys Lys Asn lie oLeu Gin Asp Ala Gly lie Asp Ala Leu Gly Lys Glu Lys Lys Asn lie o

115 120 125115 120 125

Gly Cys Val Leu Gly lie Gly Gly Gly Gin Lys Ser Ser His Glu Phe 130 135 140Gly Cys Val Leu Gly lie Gly Gly Gly Gin Lys Ser Ser His Glu Phe 130 135 140

Tyr Ser Arg Leu Asn Tyr Val Val Val Glu Lys Val Leu Arg Lys Met 145 150 155 160Tyr Ser Arg Leu Asn Tyr Val Val Val Glu Lys Val Leu Arg Lys Met 145 150 155 160

Gly Met Pro Glu Glu Asp Val Lys Val Ala Val Glu Lys Tyr Lys Ala 165 170 175Gly Met Pro Glu Glu Asp Val Lys Val Ala Val Glu Lys Tyr Lys Ala 165 170 175

Asn Phe Pro Glu Trp Arg Leu Asp Ser Phe Pro Gly Phe Leu Gly Asn 180 185 190Asn Phe Pro Glu Trp Arg Leu Asp Ser Phe Pro Gly Phe Leu Gly Asn 180 185 190

Val Thr Ala Gly Arg Cys Thr Asn Thr Phe Asn Leu Asp Gly Met Asn 195 200 205Val Thr Ala Gly Arg Cys Thr Asn Thr Phe Asn Leu Asp Gly Met Asn 195 200 205

Cys Val Val Asp Ala Ala Cys Ala Ser Ser Leu 工le Ala Val Lys Val 210 215 220Cys Val Val Asp Ala Ala Cys Ala Ser Ser Leu Ale Val Lys Val 210 215 220

Ala lie Asp Glu Leu Leu Tyr Gly Asp Cys Asp Met Met Val Thr Gly 225 230 235 240Ala lie Asp Glu Leu Leu Tyr Gly Asp Cys Asp Met Met Val Thr Gly 225 230 235 240

Ala Thr Cys Thr Asp Asn Ser lie Gly Met Tyr Met Ala Phe Ser Lys 245 250 255Ala Thr Cys Thr Asp Asn Ser lie Gly Met Tyr Met Ala Phe Ser Lys 245 250 255

Thr Pro Val Phe Ser Thr Asp Pro Ser Val Arg Ala Tyr Asp Glu Lys 260 265 270Thr Pro Val Phe Ser Thr Asp Pro Ser Val Arg Ala Tyr Asp Glu Lys 260 265 270

Thr Lys Gly Met Leu lie Gly Glu Gly Ser Ala Met Leu Val Leu Lys 275 280 285Thr Lys Gly Met Leu lie Gly Glu Gly Ser Ala Met Leu Val Leu Lys 275 280 285

Arg Tyr Ala Asp Ala Val Arg Asp Gly Asp Glu lie His Ala Val lie 290 295 300Arg Tyr Ala Asp Ala Val Arg Asp Gly Asp Glu lie His Ala Val lie 290 295 300

Arg Gly Cys Ala Ser Ser Ser Asp Gly Lys Ala Ala Gly lie Tyr Thr 305 310 315 320Arg Gly Cys Ala Ser Ser Ser Asp Gly Lys Ala Ala Gly lie Tyr Thr 305 310 315 320

Pro Thr lie Ser Gly Gin Glu Glu Ala Leu Arg Arg Ala Tyr Asn Arg 65 201038734 325 330 335Pro Thr lie Ser Gly Gin Glu Glu Ala Leu Arg Arg Ala Tyr Asn Arg 65 201038734 325 330 335

Ala Cys Val Asp Pro Ala Thr Val Thr Leu Val Glu Gly His Gly Thr 340 345 350Ala Cys Val Asp Pro Ala Thr Val Thr Leu Val Glu Gly His Gly Thr 340 345 350

a V o 5 r 5 p 3 r h T y -—I Ga V o 5 r 5 p 3 r h T y -—I G

Gly Asp Arg lie Glu· Leu Thr Ala Leu Arg Asn Leu 360 365Gly Asp Arg lie Glu· Leu Thr Ala Leu Arg Asn Leu 360 365

Phe Asp Lys Ala Tyr Gly Glu Gly Asn Thr Glu Lys Val Ala Val Gly 370 375 380Phe Asp Lys Ala Tyr Gly Glu Gly Asn Thr Glu Lys Val Ala Val Gly 370 375 380

Ser lie Lys Ser Ser 工le Gly His Leu Lys Ala Val Ala Gly Leu Ala 385 390 395 400Ser lie Lys Ser Ser Le Gly His Leu Lys Ala Val Ala Gly Leu Ala 385 390 395 400

Gly Met lie Lys Val 工le Met Ala Leu Lys His Lys Thr Leu Pro Gly 405 410 415 o r p r o h 3 T 4Gly Met lie Lys Val work le Met Ala Leu Lys His Lys Thr Leu Pro Gly 405 410 415 o r p r o h 3 T 4

Thr lie Asn Val Asp Asn Pro Pro Asn Leu Tyr Asp Asn 420 425Thr lie Asn Val Asp Asn Pro Pro Asn Leu Tyr Asp Asn 420 425

G n s A r e s r 5 e 3 s 4G n s A r e s r 5 e 3 s 4

Leu Tyr lie Asn Thr Met Asn Arg Pro Trp Phe Pro 440 445Leu Tyr lie Asn Thr Met Asn Arg Pro Trp Phe Pro 440 445

Pro Pro Gly Val Pro Arg Arg Ala 450 455Pro Pro Gly Val Pro Arg Arg Ala 450 455

Gly lie Ser Ser Phe Gly Phe Gly 460Gly lie Ser Ser Phe Gly Phe Gly 460

Gly Ala Asn Tyr His Ala Val Leu 465 470Gly Ala Asn Tyr His Ala Val Leu 465 470

Glu Glu Ala Glu Pro Glu His Thr 475 480Glu Glu Ala Glu Pro Glu His Thr 475 480

Thr Ala Tyr Arg Leu Asn Lys Arg 485Thr Ala Tyr Arg Leu Asn Lys Arg 485

Pro Gin Pro Val Leu Met Met Ala 490 495Pro Gin Pro Val Leu Met Met Ala 490 495

Ala Thr Pro Ala Ala Leu Gin Ser 500Ala Thr Pro Ala Ala Leu Gin Ser 500

Leu Cys Glu Ala Gin Leu Lys Glu 505 510Leu Cys Glu Ala Gin Leu Lys Glu 505 510

Phe Glu Ala Ala lie Lys Glu Asn 515 520Phe Glu Ala Ala lie Lys Glu Asn 515 520

Glu Thr Val Lys Asn Thr Ala Tyr 525 y' s o y3 L 5Glu Thr Val Lys Asn Thr Ala Tyr 525 y' s o y3 L 5

a Va V

r e sy I—I G 〇 r p e o h 4 p 5 s y L e h p n G1u _—t G y5 1 3 G 5 e h p s Y' L a I—_ A 〇5 r 4 p 5r e sy I—I G 〇 r p e o h 4 p 5 s y L e h p n G1u _−t G y5 1 3 G 5 e h p s Y′ L a I—_ A 〇5 r 4 p 5

Thr Asn Ala Arg Leu Gly Phe Leu Val Lys Asp Ala Glu Asp 550 555 560Thr Asn Ala Arg Leu Gly Phe Leu Val Lys Asp Ala Glu Asp 550 555 560

Ala Cys Ser Thr Leu Arg Ala lie Cys Ala Gin Phe Ala Lys Asp Val 565 570 575Ala Cys Ser Thr Leu Arg Ala lie Cys Ala Gin Phe Ala Lys Asp Val 565 570 575

Thr Lys Glu Ala Trp Arg Leu Pro Arg Glu Gly Val Ser Phe Arg Ala 580 585 590Thr Lys Glu Ala Trp Arg Leu Pro Arg Glu Gly Val Ser Phe Arg Ala 580 585 590

Lys Gly 工le Ala Thr Asn Gly Ala Val Ala Ala Leu Phe Ser Gly Gin 595 600 605Lys Gly work le Ala Thr Asn Gly Ala Val Ala Ala Leu Phe Ser Gly Gin 595 600 605

Gly Ala Gin Tyr Thr His Met Phe Ser Glu Val Ala Met Asn Trp Pro 610 615 620 66 201038734Gly Ala Gin Tyr Thr His Met Phe Ser Glu Val Ala Met Asn Trp Pro 610 615 620 66 201038734

Gin Phe Arg Gin Ser lie Ala Ala Met Asp Ala Ala Gin Ser Lys Val 625 630 635 640Gin Phe Arg Gin Ser lie Ala Ala Met Asp Ala Ala Gin Ser Lys Val 625 630 635 640

Ala Gly Ser Asp Lys Asp Phe Glu Arg Val Ser Gin Val Leu Tyr Pro 645 650 655Ala Gly Ser Asp Lys Asp Phe Glu Arg Val Ser Gin Val Leu Tyr Pro 645 650 655

Arg Lys Pro Tyr Glu Arg Glu Pro Glu Gin Asn Pro Lys Lys lie Ser 660 665 670Arg Lys Pro Tyr Glu Arg Glu Pro Glu Gin Asn Pro Lys Lys lie Ser 660 665 670

Leu Thr Ala Tyr Ser Gin Pro Ser Thr Leu Ala Cys Ala Leu Gly Ala 675 680 685Leu Thr Ala Tyr Ser Gin Pro Ser Thr Leu Ala Cys Ala Leu Gly Ala 675 680 685

Phe Glu lie Phe Lys Glu Ala Gly Phe Thr Pro Asp Phe Ala Ala Gly 690 695 700Phe Glu lie Phe Lys Glu Ala Gly Phe Thr Pro Asp Phe Ala Ala Gly 690 695 700

His Ser Leu Gly Glu Phe Ala Ala Leu Tyr Ala Ala Gly Cys Val Asp 705 710 715 720 〇His Ser Leu Gly Glu Phe Ala Ala Leu Tyr Ala Ala Gly Cys Val Asp 705 710 715 720 〇

Arg Asp Glu Leu Phe Glu Leu Val Cys Arg Arg Ala Arg lie Met Gly 725 730 735Arg Asp Glu Leu Phe Glu Leu Val Cys Arg Arg Ala Arg lie Met Gly 725 730 735

Gly Lys Asp Ala Pro Ala Thr Pro Lys Gly Cys Met Ala Ala Val lie 740 745 750Gly Lys Asp Ala Pro Ala Thr Pro Lys Gly Cys Met Ala Ala Val lie 740 745 750

Gly Pro Asn Ala Glu Asn lie Lys Val Gin Ala Ala Asn Val Trp Leu 755 760 765Gly Pro Asn Ala Glu Asn lie Lys Val Gin Ala Ala Asn Val Trp Leu 755 760 765

Gly Asn Ser Asn Ser Pro Ser Gin Thr Val lie Thr Gly Ser Val Glu 770 775 780Gly Asn Ser Asn Ser Pro Ser Gin Thr Val lie Thr Gly Ser Val Glu 770 775 780

Gly lie Gin Ala Glu Ser Ala Arg Leu Gin Lys Glu Gly Phe Arg Val 785 790 795 800Gly lie Gin Ala Glu Ser Ala Arg Leu Gin Lys Glu Gly Phe Arg Val 785 790 795 800

Val Pro Leu Ala Cys Glu Ser Ala Phe His Ser Pro Gin Met Glu Asn 805 810 815Val Pro Leu Ala Cys Glu Ser Ala Phe His Ser Pro Gin Met Glu Asn 805 810 815

Ala Ser Ser Ala Phe Lys Asp Val lie Ser Lys Val Ser Phe Arg Thr 820 825 830Ala Ser Ser Ala Phe Lys Asp Val lie Ser Lys Val Ser Phe Arg Thr 820 825 830

Pro Lys Ala Glu Thr Lys Leu Phe Ser Asn Val Ser Gly Glu Thr Tyr 835 840 845Pro Lys Ala Glu Thr Lys Leu Phe Ser Asn Val Ser Gly Glu Thr Tyr 835 840 845

Pro Thr Asp Ala Arg Glu Met Leu Thr Gin His Met Thr Ser Ser Val 850 855 860Pro Thr Asp Ala Arg Glu Met Leu Thr Gin His Met Thr Ser Ser Val 850 855 860

Lys Phe Leu Thr Gin Val Arg Asn Met His Gin Ala Gly Ala Arg lie 865 870 875 880Lys Phe Leu Thr Gin Val Arg Asn Met His Gin Ala Gly Ala Arg lie 865 870 875 880

Phe Val Glu Phe Gly Pro Lys Gin Val Leu Ser Lys Leu Val Ser Glu 885 890 895Phe Val Glu Phe Gly Pro Lys Gin Val Leu Ser Lys Leu Val Ser Glu 885 890 895

Thr Leu Lys Asp Asp Pro Ser Val Val Thr Val Ser Val Asn Pro Ala 900 905 910Thr Leu Lys Asp Asp Pro Ser Val Val Thr Val Ser Val Asn Pro Ala 900 905 910

Ser Gly Thr Asp Ser Asp lie Gin Leu Arg Asp Ala Ala Val Gin Leu 915 920 925 67 201038734Ser Gly Thr Asp Ser Asp lie Gin Leu Arg Asp Ala Ala Val Gin Leu 915 920 925 67 201038734

Val Val Ala Gly Val Asn Leu Gin Gly Phe Asp Lys Trp Asp Ala Pro 930 935 940Val Val Ala Gly Val Asn Leu Gin Gly Phe Asp Lys Trp Asp Ala Pro 930 935 940

Asp Ala Thr Arg Met Gin Ala lie Lys Lys Lys Arg Thr Thr Leu Arg 945 950 955 960Asp Ala Thr Arg Met Gin Ala lie Lys Lys Lys Arg Thr Thr Leu Arg 945 950 955 960

Leu Ser Ala Ala Thr Tyr Val Ser Asp Lys Thr Lys Lys Val Arg Asp 965 970 975Leu Ser Ala Ala Thr Tyr Val Ser Asp Lys Thr Lys Lys Val Arg Asp 965 970 975

Ala Ala Met Asn Asp Gly Arg Cys Val Thr Tyr Leu Lys Gly Ala Ala 980 985 990Ala Ala Met Asn Asp Gly Arg Cys Val Thr Tyr Leu Lys Gly Ala Ala 980 985 990

Pro Leu lie Lys Ala Pro Glu Pro Val Val Asp Glu Ala Ala Lys Arg 995 1000 1005Pro Leu lie Lys Ala Pro Glu Pro Val Val Asp Glu Ala Ala Lys Arg 995 1000 1005

Glu Ala Glu Arg Leu Gin Lys Glu Leu Gin Asp Ala Gin Arg Gin 1010 1015 1020Glu Ala Glu Arg Leu Gin Lys Glu Leu Gin Asp Ala Gin Arg Gin 1010 1015 1020

Leu Asp Asp Ala Lys Arg Ala Ala Ala Glu Ala Asn Ser Lys Leu 1025 1030 1035Leu Asp Asp Ala Lys Arg Ala Ala Ala Glu Ala Asn Ser Lys Leu 1025 1030 1035

Ala Ala Ala Lys Glu Glu Ala Lys Thr Ala Ala Ala Ser Ala Lys 1040 1045 1050Ala Ala Ala Lys Glu Glu Ala Lys Thr Ala Ala Ala Ser Ala Lys 1040 1045 1050

Pro Ala Val Asp Thr Ala Val Val Glu Lys His Arg Ala lie Leu 1055 1060 1065Pro Ala Val Asp Thr Ala Val Val Glu Lys His Arg Ala lie Leu 1055 1060 1065

Lys Ser Met Leu Ala Glu Leu Asp Gly Tyr Gly Ser Val Asp Ala 1070 1075 1080Lys Ser Met Leu Ala Glu Leu Asp Gly Tyr Gly Ser Val Asp Ala 1070 1075 1080

Ser Ser Leu Gin Gin Gin Gin Gin Gin Gin Thr Ala Pro Ala Pro 1085 1090 1095Ser Ser Leu Gin Gin Gin Gin Gin Gin Gin Thr Ala Pro Ala Pro 1085 1090 1095

Val Lys Ala Ala Ala Pro Ala Ala Pro Val Ala Ser Ala Pro Ala 1100 1105 1110Val Lys Ala Ala Ala Pro Ala Ala Pro Val Ala Ser Ala Pro Ala 1100 1105 1110

Pro Ala Val Ser Asn Glu Leu Leu Glu Lys Ala Glu Thr Val Val 1115 1120 1125Pro Ala Val Ser Asn Glu Leu Leu Glu Lys Ala Glu Thr Val Val 1115 1120 1125

Met Glu Val Leu Ala Ala Lys Thr Gly Tyr Glu Thr Asp Met lie 1130 1135 1140Met Glu Val Leu Ala Ala Lys Thr Gly Tyr Glu Thr Asp Met lie 1130 1135 1140

Glu Ala Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie 1145 1150 1155Glu Ala Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie 1145 1150 1155

Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Met Leu Asn Val 1160 1165 1170Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Met Leu Asn Val 1160 1165 1170

Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly 1175 1180 1185Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly 1175 1180 1185

Glu Val Val Asn Ala Met Lys Ala Glu lie Ala Gly Ser Ser Ala 1190 1195 1200Glu Val Val Asn Ala Met Lys Ala Glu lie Ala Gly Ser Ser Ala 1190 1195 1200

Pro Ala Pro Ala Ala Ala Ala Pro Ala Pro Ala Lys Ala Ala Pro 1205 1210 1215 68 201038734Pro Ala Pro Ala Ala Ala Ala Pro Ala Pro Ala Lys Ala Ala Pro 1205 1210 1215 68 201038734

Ala Ala 1220 Ala Ala Pro Ala Val 1225 Ser Asn Glu Leu Leu 1230 Glu Lys Ala Glu Thr 1235 Val Val Met Glu Val 1240 Leu Ala Ala Lys Thr 1245 Gly Tyr Glu Thr Asp 1250 Met lie Glu Ser Asp 1255 Met Glu Leu Glu Thr 1260 Glu Leu Gly lie Asp 1265 Ser lie Lys Arg Val 1270 Glu lie Leu Ser Glu 1275 Val Gin Ala Met Leu 1280 Asn Val Glu Ala Lys 1285 Asp Val Asp Ala Leu 1290 Ser Arg Thr ❹ Arg Thr 1295 Val Gly Glu Val Val 1300 Asn Ala Met Lys Ala 1305 Glu lie Ala Gly Gly 1310 Ser Ala Pro Ala Pro 1315 Ala Ala Ala Ala Pro 1320 Gly Pro Ala Ala Ala 1325 Ala Pro Ala Pro Ala 1330 Ala Ala Ala Pro Ala 1335 Val Ser Asn - Glu Leu 1340 Leu Glu Lys Ala Glu 1345 Thr Val Val Met Glu 1350 Val Leu Ala • Ala Lys 1355 Thr Gly Tyr Glu Thr 1360 Asp Met lie Glu Ser 1365 Asp Met Glu Leu Glu 1370 Thr Glu Leu Gly lie 1375 Asp Ser lie Lys Arg 1380 Val Glu lie Leu Ser 1385 Glu Val Gin Ala Met 1390 Leu Asn Val Glu Ala 1395 Lys Asp Val Ο Asp Ala 1400 Leu Ser Arg Thr Arg 1405 Thr Val Gly Glu Val 1410 Val Asp Ala Met Lys 1415 Ala Glu lie Ala Gly 1420 Gly Ser Ala Pro Ala 1425 Pro Ala Ala Ala Ala 1430 Pro Ala Pro Ala Ala 1435 Ala Ala Pro Ala Pro 1440 Ala Ala Pro Ala Pro 1445 Ala Val Ser Ser Glu 1450 Leu Leu Glu Lys Ala 1455 Glu Thr Val Val Met 1460 Glu Val Leu Ala Ala 1465 Lys Thr Gly Tyr Glu 1470 Thr Asp Met lie Glu 1475 Ser Asp Met Glu Leu 1480 Glu Thr Glu Leu Gly 1485 lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Met Leu Asn 69 201038734 1490 1495 1500Ala Ala 1220 Ala Ala Pro Ala Val 1225 Ser Asn Glu Leu Leu 1230 Glu Lys Ala Glu Thr 1235 Val Val Met Glu Val 1240 Leu Ala Ala Lys Thr 1245 Gly Tyr Glu Thr Asp 1250 Met lie Glu Ser Asp 1255 Met Glu Leu Glu Thr 1260 Glu Leu Gly lie Asp 1265 Ser lie Lys Arg Val 1270 Glu lie Leu Ser Glu 1275 Val Gin Ala Met Leu 1280 Asn Val Glu Ala Lys 1285 Asp Val Asp Ala Leu 1290 Ser Arg Thr ❹ Arg Thr 1295 Val Gly Glu Val Val 1300 Asn Ala Met Lys Ala 1305 Glu lie Ala Gly Gly 1310 Ser Ala Pro Ala Pro 1315 Ala Ala Ala Ala Pro 1320 Gly Pro Ala Ala Ala 1325 Ala Pro Ala Pro Ala 1330 Ala Ala Ala Pro Ala 1335 Val Ser Asn - Glu Leu 1340 Leu Glu Lys Ala Glu 1345 Thr Val Val Met Glu 1350 Val Leu Ala • Ala Lys 1355 Thr Gly Tyr Glu Thr 1360 Asp Met lie Glu Ser 1365 Asp Met Glu Leu Glu 1370 Thr Glu Leu Gly lie 1375 Asp Ser lie Lys Arg 1380 Val Glu Lie Leu Ser 1385 Glu Val Gin Ala Met 1390 Leu Asn Val Glu Ala 1395 Lys Asp Val Ο Asp Ala 1400 Leu Ser Arg Thr Arg 1405 Thr Val Gly Glu Val 1410 Val Asp Ala Met Lys 1415 Ala Glu lie Ala Gly 1420 Gly Ser Ala Pro Ala 1425 Pro Ala Ala Ala Ala 1430 Pro Ala Pro Ala Ala 1435 Ala Ala Pro Ala Pro 1440 Ala Ala Pro Ala Pro 1445 Ala Val Ser Ser Glu 1450 Leu Leu Glu Lys Ala 1455 Glu Thr Val Val Met 1460 Glu Val Leu Ala Ala 1465 Lys Thr Gly Tyr Glu 1470 Thr Asp Met lie Glu 1475 Ser Asp Met Glu Leu 1480 Glu Thr Glu Leu Gly 1485 lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Met Leu Asn 69 201038734 1490 1495 1500

Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val 1505 1510 1515Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val 1505 1510 1515

Gly Glu Val Val Asp Ala Met Lys Ala Glu lie Ala Gly Gly Ser 1520 1525 1530Gly Glu Val Val Asp Ala Met Lys Ala Glu lie Ala Gly Gly Ser 1520 1525 1530

Ala Pro Ala Pro Ala Ala Ala Ala Pro Ala Pro Ala Ala Ala Ala 1535 1540 1545Ala Pro Ala Pro Ala Ala Ala Ala Pro Ala Pro Ala Ala Ala Ala 1535 1540 1545

Pro Ala Pro Ala Ala Pro Ala Pro Ala Ala Pro Ala Pro Ala Val 1550 1555 1560Pro Ala Pro Ala Ala Pro Ala Pro Ala Ala Pro Ala Pro Ala Val 1550 1555 1560

Ser Ser Glu Leu Leu Glu Lys Ala Glu Thr Val Val Met Glu Val 1565 1570 1575Ser Ser Glu Leu Leu Glu Lys Ala Glu Thr Val Val Met Glu Val 1565 1570 1575

Leu Ala Ala Lys Thr Gly Tyr Glu Thr Asp Met lie Glu Ser Asp 1580 1585 1590Leu Ala Ala Lys Thr Gly Tyr Glu Thr Asp Met lie Glu Ser Asp 1580 1585 1590

Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val 1595 1600 1605Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val 1595 1600 1605

Glu lie Leu Ser Glu Val Gin Ala Met Leu Asn Val Glu Ala Lys 1610 1615 1620Glu lie Leu Ser Glu Val Gin Ala Met Leu Asn Val Glu Ala Lys 1610 1615 1620

Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val 1625 1630 1635Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val 1625 1630 1635

Asp Ala Met Lys Ala Glu lie Ala Gly Ser Ser Ala Ser Ala Pro 1640 1645 1650Asp Ala Met Lys Ala Glu lie Ala Gly Ser Ser Ala Ser Ala Pro 1640 1645 1650

Ala Ala Ala Ala Pro Ala Pro Ala Ala Ala Ala Pro Ala Pro Ala 1655 1660 1665Ala Ala Ala Ala Pro Ala Pro Ala Ala Ala Ala Pro Ala Pro Ala 1655 1660 1665

Ala Ala Ala Pro Ala Val Ser Asn Glu Leu Leu Glu Lys Ala Glu 1670 1675 1680Ala Ala Ala Pro Ala Val Ser Asn Glu Leu Leu Glu Lys Ala Glu 1670 1675 1680

Thr Val Val Met Glu Val Leu Ala Ala Lys Thr Gly Tyr Glu Thr 1685 1690 1695Thr Val Val Met Glu Val Leu Ala Ala Lys Thr Gly Tyr Glu Thr 1685 1690 1695

Asp Met lie Glu Ser Asp Met Glu Leu Glu Thr Glu Leu Gly lie 1700 1705 1710Asp Met lie Glu Ser Asp Met Glu Leu Glu Thr Glu Leu Gly lie 1700 1705 1710

Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Met 1715 1720 1725Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Met 1715 1720 1725

Leu Asn Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg 1730 1735 1740Leu Asn Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg 1730 1735 1740

Thr Val Gly Glu Val Val Asp Ala Met Lys Ala Glu lie Ala Gly 1745 1750 1755Thr Val Gly Glu Val Val Asp Ala Met Lys Ala Glu lie Ala Gly 1745 1750 1755

Gly Ser Ala Pro Ala Pro Ala Ala Ala Ala Pro Ala Pro Ala Ala 1760 1765 1770 70 201038734Gly Ser Ala Pro Ala Pro Ala Ala Ala Ala Pro Ala Pro Ala Ala 1760 1765 1770 70 201038734

Ala Ala Pro Ala Val Ser Asn Glu Leu Leu Glu Lys Ala Glu Thr 1775 1780 1785Ala Ala Pro Ala Val Ser Asn Glu Leu Leu Glu Lys Ala Glu Thr 1775 1780 1785

Val Val Met Glu Val Leu Ala Ala Lys Thr Gly Tyr Glu Thr Asp 1790 1795 1800Val Val Met Glu Val Leu Ala Ala Lys Thr Gly Tyr Glu Thr Asp 1790 1795 1800

Met lie Glu Ser Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp 1805 1810 1815Met lie Glu Ser Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp 1805 1810 1815

Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Met Leu 1820 1825 1830Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Met Leu 1820 1825 1830

Asn Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr 1835 1840 1845Asn Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr 1835 1840 1845

Val Gly Glu Val Val Asp Ala Met Lys Ala Glu lie Ala Gly Ser 1850 1855 1860 ΟVal Gly Glu Val Val Asp Ala Met Lys Ala Glu lie Ala Gly Ser 1850 1855 1860 Ο

Ser Ala Pro Ala Pro Ala Ala Ala Ala Pro Ala Pro Ala Ala Ala 1865 1870 1875Ser Ala Pro Ala Pro Ala Ala Ala Ala Pro Ala Pro Ala Ala Ala 1865 1870 1875

Ala Pro Ala Pro Ala Ala Ala Ala Pro Ala Val Ser Ser Glu Leu 1880 1885 1890Ala Pro Ala Pro Ala Ala Ala Ala Ala Val Ser Ser Glu Leu 1880 1885 1890

Leu Glu Lys Ala Glu Thr Val Val Met Glu Val Leu Ala Ala Lys 1895 1900 1905Leu Glu Lys Ala Glu Thr Val Val Met Glu Val Leu Ala Ala Lys 1895 1900 1905

Thr Gly Tyr Glu Thr Asp Met lie Glu Ser Asp Met Glu Leu Glu 1910 1915 1920Thr Gly Tyr Glu Thr Asp Met lie Glu Ser Asp Met Glu Leu Glu 1910 1915 1920

Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser 1925 1930 1935Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser 1925 1930 1935

Glu Val Gin Ala Met Leu Asn Val Glu Ala Lys Asp Val Asp Ala 1940 1945 1950Glu Val Gin Ala Met Leu Asn Val Glu Ala Lys Asp Val Asp Ala 1940 1945 1950

Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met Lys 1955 I960 1965Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met Lys 1955 I960 1965

Ala Glu lie Ala Gly Gly Ser Ala Pro Ala Pro Ala Ala Ala Ala 1970 1975 1980Ala Glu lie Ala Gly Gly Ser Ala Pro Ala Pro Ala Ala Ala Ala 1970 1975 1980

Pro Ala Pro Ala Ala Ala Ala Pro Ala Val Ser Asn Glu Leu Leu 1985 1990 1995Pro Ala Pro Ala Ala Ala Ala Pro Ala Val Ser Asn Glu Leu Leu 1985 1990 1995

Glu Lys Ala Glu Thr Val Val Met Glu Val Leu Ala Ala Lys Thr 2000 2005 2010Glu Lys Ala Glu Thr Val Val Met Glu Val Leu Ala Ala Lys Thr 2000 2005 2010

Gly Tyr Glu Thr Asp Met lie Glu Ser Asp Met Glu Leu Glu Thr 2015 2020 2025Gly Tyr Glu Thr Asp Met lie Glu Ser Asp Met Glu Leu Glu Thr 2015 2020 2025

Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu 2030 2035 2040Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu 2030 2035 2040

Val Gin Ala Met Leu Asn Val Glu Ala Lys Asp Val Asp Ala Leu 2045 2050 2055 71 201038734Val Gin Ala Met Leu Asn Val Glu Ala Lys Asp Val Asp Ala Leu 2045 2050 2055 71 201038734

Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala 2060 2065 2070Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala 2060 2065 2070

Glu lie Ala Gly Gly Ser Ala Pro Ala Pro Ala Ala Ala Ala Pro 2075 2080 2085Glu lie Ala Gly Gly Ser Ala Pro Ala Pro Ala Ala Ala Ala Pro 2075 2080 2085

Ala Ser Ala Gly Ala Ala Pro Ala Val Lys lie Asp Ser Val His 2090 2095 2100Ala Ser Ala Gly Ala Ala Pro Ala Val Lys lie Asp Ser Val His 2090 2095 2100

Gly Ala Asp Cys Asp Asp Leu Ser Leu Met His Ala Lys Val Val 2105 2110 2115Gly Ala Asp Cys Asp Asp Leu Ser Leu Met His Ala Lys Val Val 2105 2110 2115

Asp lie Arg Arg Pro Asp Glu Leu lie Leu Glu Arg Pro Glu Asn 2120 2125 2130Asp lie Arg Arg Pro Asp Glu Leu lie Leu Glu Arg Pro Glu Asn 2120 2125 2130

Arg Pro Val Leu Val Val Asp Asp Gly Ser Glu Leu Thr Leu Ala 2135 2140 2145Arg Pro Val Leu Val Val Asp Asp Gly Ser Glu Leu Thr Leu Ala 2135 2140 2145

OO

Leu Val Arg Val Leu Gly Ala Cys Ala Val Val Leu Thr Phe Glu 2150 2155 2160Leu Val Arg Val Leu Gly Ala Cys Ala Val Val Leu Thr Phe Glu 2150 2155 2160

Gly Leu Gin Leu Ala Gin Arg Ala Gly Ala Ala Ala lie Arg His 2165 2170 2175Gly Leu Gin Leu Ala Gin Arg Ala Gly Ala Ala Ala lie Arg His 2165 2170 2175

Lys Ala lie Lys Glu Ala Glu Gin Arg Phe Gly Ala Leu Gly Gly 2180 2185 2190Lys Ala lie Lys Glu Ala Glu Gin Arg Phe Gly Ala Leu Gly Gly 2180 2185 2190

Phe He Ser Gin Gin Ala Glu Arg Phe Glu Pro Ala Glu lie Leu 2195 2200 2205Phe He Ser Gin Gin Ala Glu Arg Phe Glu Pro Ala Glu lie Leu 2195 2200 2205

Gly Phe Thr Leu Met Cys Ala Lys Phe Ala Lys Ala Ser Leu Cys 2210 2215 2220Gly Phe Thr Leu Met Cys Ala Lys Phe Ala Lys Ala Ser Leu Cys 2210 2215 2220

Thr Ala Val Ala Gly Val Leu Ala Lys Asp Leu Ser Ala Glu Ser 2225 2230 2235 ❹Thr Ala Val Ala Gly Val Leu Ala Lys Asp Leu Ser Ala Glu Ser 2225 2230 2235 ❹

Ala Glu Gly Arg Pro Ala Phe lie Gly Val Ala Arg Leu Asp Gly 2240 2245 2250Ala Glu Gly Arg Pro Ala Phe lie Gly Val Ala Arg Leu Asp Gly 2240 2245 2250

Arg Leu Gly Phe Thr Ser Gin Gly Thr Ser Asp Ala Leu Lys Arg 2255 2260 2265Arg Leu Gly Phe Thr Ser Gin Gly Thr Ser Asp Ala Leu Lys Arg 2255 2260 2265

Ala Gin Arg Gly Ala lie Phe Gly Leu Cys Lys Thr 工le Gly Leu 2270 2275 2280Ala Gin Arg Gly Ala lie Phe Gly Leu Cys Lys Thr work le Gly Leu 2270 2275 2280

Glu Trp Ser Glu Ser Asp Val Phe Ser Arg Gly Val Asp lie Ala 2285 2290 2295Glu Trp Ser Glu Ser Asp Val Phe Ser Arg Gly Val Asp lie Ala 2285 2290 2295

Gin Gly Met His Pro Glu Asp Ala Ala Val Ala 2300 2305 a V o el 13 I 2Gin Gly Met His Pro Glu Asp Ala Ala Val Ala 2300 2305 a V o el 13 I 2

u I—I G g r Au I—I G g r A

Met Ala Cys Ala Asp lie Arg lie Arg Glu Val Gly lie Gly Ala 2315 2320 2325Met Ala Cys Ala Asp lie Arg lie Arg Glu Val Gly lie Gly Ala 2315 2320 2325

Asn Gin Gin Arg Cys Thr lie Arg Ala Ala Lys Leu Glu Thr Gly 2330 2335 2340 72 201038734Asn Gin Gin Arg Cys Thr lie Arg Ala Ala Lys Leu Glu Thr Gly 2330 2335 2340 72 201038734

Asn Pro Gin Arg Gin lie Ala Lys Asp Asp Val Leu Leu Val Ser 2345 2350 2355Asn Pro Gin Arg Gin lie Ala Lys Asp Asp Val Leu Leu Val Ser 2345 2350 2355

Gly Gly Ala Arg Gly lie Thr Pro Leu Cys lie Arg Glu lie Thr 2360 2365 2370Gly Gly Ala Arg Gly lie Thr Pro Leu Cys lie Arg Glu lie Thr 2360 2365 2370

Arg Gin 工le Ala Gly Gly Lys Tyr lie Leu Leu Gly Arg Ser Lys 2375 2380 2385Arg Gin work le Ala Gly Gly Lys Tyr lie Leu Leu Gly Arg Ser Lys 2375 2380 2385

Val Ser Ala Ser Glu Pro Ala Trp Cys Ala Gly lie Thr Asp Glu 2390 2395 2400Val Ser Ala Ser Glu Pro Ala Trp Cys Ala Gly lie Thr Asp Glu 2390 2395 2400

Lys Ala Val Gin Lys Ala Ala Thr Gin Glu Leu Lys Arg Ala Phe 2405 2410 2415 ΟLys Ala Val Gin Lys Ala Ala Thr Gin Glu Leu Lys Arg Ala Phe 2405 2410 2415 Ο

Ser Ala Gly Glu Gly Pro Lys Pro Thr Pro Arg Ala Val Thr Lys 2420 2425 2430Ser Ala Gly Glu Gly Pro Lys Pro Thr Pro Arg Ala Val Thr Lys 2420 2425 2430

Leu Val Gly Ser Val Leu Gly Ala Arg Glu Val Arg Ser Ser lie 2435 2440 2445Leu Val Gly Ser Val Leu Gly Ala Arg Glu Val Arg Ser Ser lie 2435 2440 2445

Ala Ala lie Glu Ala Leu Gly Gly Lys Ala 工le Tyr Ser Ser Cys 2450 2455 2460Ala Ala lie Glu Ala Leu Gly Gly Lys Ala l Tyr Ser Ser Cys 2450 2455 2460

Asp Val Asn Ser Ala Ala Asp Val Ala Lys Ala Val Arg Asp Ala 2465 2470 2475Asp Val Asn Ser Ala Ala Asp Val Ala Lys Ala Val Arg Asp Ala 2465 2470 2475

Glu Ser Gin Leu Gly Ala Arg Val Ser Gly lie Val His Ala Ser 2480 2485 2490Glu Ser Gin Leu Gly Ala Arg Val Ser Gly lie Val His Ala Ser 2480 2485 2490

Gly Val Leu Arg Asp Arg Leu lie Glu Lys Lys Leu Pro Asp Glu 2495 2500 2505Gly Val Leu Arg Asp Arg Leu lie Glu Lys Lys Leu Pro Asp Glu 2495 2500 2505

Phe Asp Ala Val Phe Gly Thr Lys Val Thr Gly Leu Glu Asn Leu 2510 2515 2520Phe Asp Ala Val Phe Gly Thr Lys Val Thr Gly Leu Glu Asn Leu 2510 2515 2520

Leu Ala Ala Val Asp Arg Ala Asn Leu Lys His Met Val Leu Phe 2525 2530 2535Leu Ala Ala Val Asp Arg Ala Asn Leu Lys His Met Val Leu Phe 2525 2530 2535

Ser Ser Leu Ala Gly Phe His Gly Asn Val Gly Gin Ser Asp Tyr 2540 2545 2550Ser Ser Leu Ala Gly Phe His Gly Asn Val Gly Gin Ser Asp Tyr 2540 2545 2550

Ala Met Ala Asn Glu Ala Leu Asn Lys Met Gly Leu Glu Leu Ala 2555 2560 2565Ala Met Ala Asn Glu Ala Leu Asn Lys Met Gly Leu Glu Leu Ala 2555 2560 2565

Lys Asp Val Ser Val Lys Ser lie Cys Phe Gly Pro Trp Asp Gly 2570 2575 2580Lys Asp Val Ser Val Lys Ser lie Cys Phe Gly Pro Trp Asp Gly 2570 2575 2580

Gly Met Val Thr Pro Gin Leu Lys Lys Gin Phe Gin Glu Met Gly 2585 2590 2595Gly Met Val Thr Pro Gin Leu Lys Lys Gin Phe Gin Glu Met Gly 2585 2590 2595

Val Gin lie lie Pro Arg Glu Gly Gly Ala Asp Thr Val Ala Arg 2600 2605 2610 lie Val Leu Gly Ser Ser Pro Ala Glu lie Leu Val Gly Asn Trp 73 201038734 2615 2620 2625Val Gin lie lie Pro Arg Glu Gly Gly Ala Asp Thr Val Ala Arg 2600 2605 2610 lie Val Leu Gly Ser Ser Pro Ala Glu lie Leu Val Gly Asn Trp 73 201038734 2615 2620 2625

Arg Thr Pro Ser Lys Lys Val Gly Ser Asp Thr Tie Thr Leu His 2630 2635 2640Arg Thr Pro Ser Lys Lys Val Gly Ser Asp Thr Tie Thr Leu His 2630 2635 2640

Arg Lys lie Ser Ala Lys Ser Asn Pro Phe Leu Glu Asp His Val 2645 2650 2655 lie Gin Gly Arg Arg Val Leu Pro Met Thr Leu Ala lie Gly Ser 2660 2665 2670Arg Lys lie Ser Ala Lys Ser Asn Pro Phe Leu Glu Asp His Val 2645 2650 2655 lie Gin Gly Arg Arg Val Leu Pro Met Thr Leu Ala lie Gly Ser 2660 2665 2670

Leu Ala Glu Thr Cys Leu Gly Leu Phe Pro Gly Tyr Ser Leu Trp 2675 2680 2685Leu Ala Glu Thr Cys Leu Gly Leu Phe Pro Gly Tyr Ser Leu Trp 2675 2680 2685

Ala lie Asp Asp Ala Gin Leu Phe Lys Gly Val Thr Val Asp Gly 2690 2695 2700Ala lie Asp Asp Ala Gin Leu Phe Lys Gly Val Thr Val Asp Gly 2690 2695 2700

Asp Val Asn Cys Glu Val Thr Leu Thr Pro Ser Thr Ala Pro SerAsp Val Asn Cys Glu Val Thr Leu Thr Pro Ser Thr Ala Pro Ser

2705 2710 27152705 2710 2715

Gly Arg Val Asn Val Gin Ala Thr Leu Lys Thr Phe Ser Ser Gly 2720 2725 2730Gly Arg Val Asn Val Gin Ala Thr Leu Lys Thr Phe Ser Ser Gly 2720 2725 2730

Lys Leu Val Pro Ala Tyr Arg Ala Val lie Val Leu Ser Asn Gin 2735 2740 2745Lys Leu Val Pro Ala Tyr Arg Ala Val lie Val Leu Ser Asn Gin 2735 2740 2745

Gly Ala Pro Pro Ala Asn Ala Thr Met Gin Pro Pro Ser Leu Asp 2750 2755 2760Gly Ala Pro Pro Ala Asn Ala Thr Met Gin Pro Pro Ser Leu Asp 2750 2755 2760

Ala Asp Pro Ala Leu Gin Gly Ser Val Tyr Asp Gly Lys Thr Leu 2765 2770 2775Ala Asp Pro Ala Leu Gin Gly Ser Val Tyr Asp Gly Lys Thr Leu 2765 2770 2775

Phe His Gly Pro Ala Phe Arg Gly lie Asp Asp Val Leu Ser Cys 2780 2785 2790Phe His Gly Pro Ala Phe Arg Gly lie Asp Asp Val Leu Ser Cys 2780 2785 2790

Thr Lys Ser Gin Leu Val Ala Lys Cys Ser Ala Val Pro Gly Ser 2795 2800 2805Thr Lys Ser Gin Leu Val Ala Lys Cys Ser Ala Val Pro Gly Ser 2795 2800 2805

Asp Ala Ala Arg Gly Glu Phe Ala Thr Asp Thr Asp Ala His Asp 2810 2815 2820Asp Ala Ala Arg Gly Glu Phe Ala Thr Asp Thr Asp Ala His Asp 2810 2815 2820

Pro Phe Val Asn Asp Leu Ala Phe Gin Ala Met Leu Val Trp Val 2825 2830 2835Pro Phe Val Asn Asp Leu Ala Phe Gin Ala Met Leu Val Trp Val 2825 2830 2835

Arg Arg Thr Leu Gly Gin Ala Ala Leu Pro Asn Ser lie Gin Arg 2840 2845 2850 lie Val Gin His Arg Pro Val Pro Gin Asp Lys Pro Phe Tyr lie 2855 2860 2865Arg Arg Thr Leu Gly Gin Ala Ala Leu Pro Asn Ser lie Gin Arg 2840 2845 2850 lie Val Gin His Arg Pro Val Pro Gin Asp Lys Pro Phe Tyr lie 2855 2860 2865

Thr Leu Arg Ser Asn Gin Ser Gly Gly His Ser Gin His Lys His 2870 2875 2880Thr Leu Arg Ser Asn Gin Ser Gly Gly His Ser Gin His Lys His 2870 2875 2880

Ala Leu Gin Phe His Asn Glu Gin Gly Asp Leu Phe lie Asp Val 2885 2890 2895 74 201038734Ala Leu Gin Phe His Asn Glu Gin Gly Asp Leu Phe lie Asp Val 2885 2890 2895 74 201038734

Gin Ala Ser Val He Ala Thr Asp Ser Leu Ala Phe 2900 2905 2910 <210〉 55 <211> 2395 <212> PRT <213> 金黃色破囊壺菌 <400> 55Gin Ala Ser Val A A A A A A A A A A A A A A A A A A A

Arg Lys Cys lie Arg Pro Ser Leu Gly His His Trp Ala lie lie Gly 1 5 10 15Arg Lys Cys lie Arg Pro Ser Leu Gly His His Trp Ala lie lie Gly 1 5 10 15

Val Leu Gly Arg Ala Leu Arg He Val Arg Pro He Arg Tyr Glu Ala 20 25 30Val Leu Gly Arg Ala Leu Arg He Val Arg Pro He Arg Tyr Glu Ala 20 25 30

Thr Asn Leu Arg Arg Leu Pro Arg Ser Gly Trp Leu Val Ala Leu Glv 35 40 45Thr Asn Leu Arg Arg Leu Pro Arg Ser Gly Trp Leu Val Ala Leu Glv 35 40 45

Leu Phe Cys Asp Leu Ser Ser Cys Ma Gly Lys Leu Asp Leu Gin ThrLeu Phe Cys Asp Leu Ser Ser Cys Ma Gly Lys Leu Asp Leu Gin Thr

Arg Asp Thr Ala Lys Asp Pro Cys Cys Lys Arg Lys Trp Ser Ma SerArg Asp Thr Ala Lys Asp Pro Cys Cys Lys Arg Lys Trp Ser Ma Ser

Arg Ala Pro Pro Arg Pro Arg Ala Glu Ala Asp LyS Ma Ser Asn GluArg Ala Pro Pro Arg Pro Arg Ala Glu Ala Asp LyS Ma Ser Asn Glu

Met Glu Thr Lys Asp Asp Arg Val Ala He Val Gly Met Ser Ala He 100 105 110Met Glu Thr Lys Asp Asp Arg Val Ala He Val Gly Met Ser Ala He 100 105 110

Leu Pro Cys Gly Glu Ser Val Arg Glu Ser Trp Glu Ala lie Arg Glu 115 120 125Leu Pro Cys Gly Glu Ser Val Arg Glu Ser Trp Glu Ala lie Arg Glu 115 120 125

Gly Leu Asp Cys Leu Gin Asp Leu Pro Ala Asp Arq Val Asp lie Thr 130 135 140Gly Leu Asp Cys Leu Gin Asp Leu Pro Ala Asp Arq Val Asp lie Thr 130 135 140

Ala Tyr Tyr Asp Pro Asn Lys Thr Thr Lys Asp Lys lie Tyr Cys Lys 145 150 155 160Ala Tyr Tyr Asp Pro Asn Lys Thr Thr Lys Asp Lys lie Tyr Cys Lys 145 150 155 160

Arg Gly Gly Phe lie Pro Glu Tyr Asp Phe Asp Ala Arg Glu Phe Gly 165 170 175Arg Gly Gly Phe lie Pro Glu Tyr Asp Phe Asp Ala Arg Glu Phe Gly 165 170 175

Leu Asn Met Phe Gin Met Glu Asp Ser Asp Ala Asn Gin Thr Val Thr 180 185 190Leu Asn Met Phe Gin Met Glu Asp Ser Asp Ala Asn Gin Thr Val Thr 180 185 190

Leu Leu Lys Val Lys Glu Ala Leu Glu Asp Ala Gly Val Glu Pro Phe 195 200 205Leu Leu Lys Val Lys Glu Ala Leu Glu Asp Ala Gly Val Glu Pro Phe 195 200 205

Thr Lys Lys Lys Lys Asn lie Gly Cys Val Leu Gly 工le Gly Gly Gly 210 215 220Thr Lys Lys Lys Lys Asn lie Gly Cys Val Leu Gly work le Gly Gly Gly 210 215 220

Gin Lys Ala Ser His Glu Phe Tyr Ser Arg Leu Asn Tyr Val Val Val 225 230 235 240Gin Lys Ala Ser His Glu Phe Tyr Ser Arg Leu Asn Tyr Val Val Val 225 230 235 240

Glu Lys Val Leu Arg Lys Met Asn Leu Pro Asp Glu Val Val Glu Ala 245 250 255 75 201038734Glu Lys Val Leu Arg Lys Met Asn Leu Pro Asp Glu Val Val Glu Ala 245 250 255 75 201038734

Ala Val Glu Lys Tyr Lys Ala Asn Phe Pro Glu Trp Arg Leu Asp Ser 260 265 270Ala Val Glu Lys Tyr Lys Ala Asn Phe Pro Glu Trp Arg Leu Asp Ser 260 265 270

Phe Pro Gly Phe Leu Gly Asn Val Thr Ala Gly Arg Cys Ser Asn Val 275 280 285Phe Pro Gly Phe Leu Gly Asn Val Thr Ala Gly Arg Cys Ser Asn Val 275 280 285

Phe Asn Met Glu Gly Met Asn Cys Val Val Asp Ala Ala Cys Ala Ser 290 295 300Phe Asn Met Glu Gly Met Asn Cys Val Val Asp Ala Ala Cys Ala Ser 290 295 300

Ser Leu lie Ala lie Lys Val Ala 工le Asp Glu Leu Leu His Gly Asp 305 310 315 320Ser Leu lie Ala lie Lys Val Ala gong le Asp Glu Leu Leu His Gly Asp 305 310 315 320

Cys Asp Thr Met lie Ala Gly Ala Thr Cys Thr Asp Asn Ser lie Gly 325 330 335Cys Asp Thr Met lie Ala Gly Ala Thr Cys Thr Asp Asn Ser lie Gly 325 330 335

Met Tyr Met Ala Phe Ser Lys Thr Pro Val Phe Ser Thr Asp Gin Ser 340 345 350Met Tyr Met Ala Phe Ser Lys Thr Pro Val Phe Ser Thr Asp Gin Ser 340 345 350

Val Lys Ala Tyr Asp Ala Lys Thr Lys Gly Met Leu lie Gly G丄u Gly 355 360 365Val Lys Ala Tyr Asp Ala Lys Thr Lys Gly Met Leu lie Gly G丄u Gly 355 360 365

Ser Ala Met Val Val Leu Lys Arg Tyr Ala Asp Ala Val Arg Asp Gly 370 375 380Ser Ala Met Val Val Leu Lys Arg Tyr Ala Asp Ala Val Arg Asp Gly 370 375 380

Asp Glu lie His Ala Val lie Arg Ala Cys Ala Ser Ser Ser Asp Gly 385 390 395 400Asp Glu lie His Ala Val lie Arg Ala Cys Ala Ser Ser Ser Asp Gly 385 390 395 400

Lys Ala Ala Gly lie Tyr Ala Pro Thr Val Ser Gly Gin Glu Glu Ala 405 410 415Lys Ala Ala Gly lie Tyr Ala Pro Thr Val Ser Gly Gin Glu Glu Ala 405 410 415

Leu Arg Arg Ala Tyr Ala Arg Ala Gly Val Asp Pro Ser Thr Val Thr 420 425 430Leu Arg Arg Ala Tyr Ala Arg Ala Gly Val Asp Pro Ser Thr Val Thr 420 425 430

Leu Val Glu Gly His Gly Thr Gly Thr Pro Val Gly Asp Arg 工le Glu 435 440 445Leu Val Glu Gly His Gly Thr Gly Thr Pro Val Gly Asp Arg work le Glu 435 440 445

Leu Thr Ala Leu Arg Asn Val Phe Asp Ala Ala Asn Lys Gly Arg Lys 450 455 460Leu Thr Ala Leu Arg Asn Val Phe Asp Ala Ala Asn Lys Gly Arg Lys 450 455 460

Glu Thr Val Ala Val Gly Ser lie Lys Ser Gin lie Gly His Leu Lys 465 470 475 480Glu Thr Val Ala Val Gly Ser lie Lys Ser Gin lie Gly His Leu Lys 465 470 475 480

Ala Val Ala Gly Phe Ala Gly Leu Val Lys Val Val Met Ala Leu Lys 485 490 495Ala Val Ala Gly Phe Ala Gly Leu Val Lys Val Val Met Ala Leu Lys 485 490 495

His Lys Thr Leu Pro Gin Thr lie Asn Val His Asp Pro Pro Ala Leu 500 505 510His Lys Thr Leu Pro Gin Thr lie Asn Val His Asp Pro Pro Ala Leu 500 505 510

His Asp Gly Ser Pro lie Gin Asp Ser Ser Leu Tyr lie Asn Thr Met 515 520 525His Asp Gly Ser Pro lie Gin Asp Ser Ser Leu Tyr lie Asn Thr Met 515 520 525

Asn Arg Pro Trp Phe Thr Ala Pro Gly Val Pro Arg Arg Ala Gly lie 530 535 540Asn Arg Pro Trp Phe Thr Ala Pro Gly Val Pro Arg Arg Ala Gly lie 530 535 540

Ser Ser Phe Gly Phe Gly Gly Ala Asn Tyr His Ala Val Leu Glu Glu 545 550 555 560 76 201038734Ser Ser Phe Gly Phe Gly Gly Ala Asn Tyr His Ala Val Leu Glu Glu 545 550 555 560 76 201038734

Ala Glu Pro Glu His Ala Lys Pro Tyr Arg Met Asn Gin Val Pro Gin 565 570 575 a V o r pAla Glu Pro Glu His Ala Lys Pro Tyr Arg Met Asn Gin Val Pro Gin 565 570 575 a V o r p

s Y' c e _—_ I r o e 9 s 5 a 1 A u e L a _—_ A r e s a 5 18 A 5 r e s r e s a _—I A s i H uo e 8 L 5 u e Ls Y' c e ___ I r o e 9 s 5 a 1 A u e L a ___ A r e s a 5 18 A 5 r e s r e s a _—I A s i H uo e 8 L 5 u e L

Asp Ala Gin Ala Asp Ala Leu Gin Ala Ala Val Ser Pro Glu Ala Ser 595 600 605Asp Ala Gin Ala Asp Ala Leu Gin Ala Ala Val Ser Pro Glu Ala Ser 595 600 605

Lys His Ala Asp Tyr Arg Ala lie Val Ala Phe His Glu Ala Phe Lys 610 615 620Lys His Ala Asp Tyr Arg Ala lie Val Ala Phe His Glu Ala Phe Lys 610 615 620

Leu Arg Ala Gly Val Pro Ala Gly His Ala Arg lie Gly Phe Val Ser 625 630 635 640Leu Arg Ala Gly Val Pro Ala Gly His Ala Arg lie Gly Phe Val Ser 625 630 635 640

Gly Ser Ala Ala Ala Thr Leu Ala Val Leu Arg Ala Ala Ser Ala Lys Ο o 645 650 655Gly Ser Ala Ala Ala Thr Leu Ala Val Leu Arg Ala Ala Ser Ala Lys Ο o 645 650 655

Leu Lys Gin Ser Ser Ala Thr Leu Glu Trp Thr Leu Leu Arg Glu Gly 660 665 67 0Leu Lys Gin Ser Ser Ala Thr Leu Glu Trp Thr Leu Leu Arg Glu Gly 660 665 67 0

Val Thr Tyr Arg Ser Ala Ala Met His Thr Pro Gly Ser Val Ala Ala 675 680 685Val Thr Tyr Arg Ser Ala Ala Met His Thr Pro Gly Ser Val Ala Ala 675 680 685

Leu Phe Ala Gly Gin Gly Ala Gin Tyr Thr His Met Phe Ala Asp Val 690 695 700Leu Phe Ala Gly Gin Gly Ala Gin Tyr Thr His Met Phe Ala Asp Val 690 695 700

Ala Met Asn Trp Pro Pro Phe Arg Ser Ala Val Gin Glu Met Asp Ala 705 710 715 720Ala Met Asn Trp Pro Pro Phe Arg Ser Ala Val Gin Glu Met Asp Ala 705 710 715 720

Ala Gin Val Thr Ala Ala Ala Pro Lys Arg Leu Ser Glu Val Leu Tyr 725 730 735Ala Gin Val Thr Ala Ala Ala Pro Lys Arg Leu Ser Glu Val Leu Tyr 725 730 735

Pro Arg Lys Pro Tyr Ala Ala Glu Pro Glu Gin Asp Asn Lys Ala lie 740 745 750Pro Arg Lys Pro Tyr Ala Ala Glu Pro Glu Gin Asp Asn Lys Ala lie 740 745 750

Ser Met Thr lie Asn Ser Gin Pro Ala Leu Met Ala Cys Ala Ala Gly 755 760 765Ser Met Thr lie Asn Ser Gin Pro Ala Leu Met Ala Cys Ala Ala Gly 755 760 765

Ala Phe Glu Val Phe Arg Gin Ala Gly Leu Ala Pro Asp His Val Ala 770 775 780Ala Phe Glu Val Phe Arg Gin Ala Gly Leu Ala Pro Asp His Val Ala 770 775 780

Gly His Ser Leu Gly Glu Phe Gly Ala Leu Leu Ala Ala Gly Cys Ala 785 790 795 800Gly His Ser Leu Gly Glu Phe Gly Ala Leu Leu Ala Ala Gly Cys Ala 785 790 795 800

Ser Arg Glu Glu Leu Phe Arg Leu Val Cys Ser Arg Ala Lys Ala Met 805 810 815Ser Arg Glu Glu Leu Phe Arg Leu Val Cys Ser Arg Ala Lys Ala Met 805 810 815

Gin Asp Val Pro Gin Gly Asp Gly Ala Trp Leu Ala Asn Cys Asn Ser 820 825 830Gin Asp Val Pro Gin Gly Asp Gly Ala Trp Leu Ala Asn Cys Asn Ser 820 825 830

Pro Ser Gin Val Val lie Ser Gly Asp Lys Thr Ala Val Glu Arg Glu 835 840 845Pro Ser Gin Val Val lie Ser Gly Asp Lys Thr Ala Val Glu Arg Glu 835 840 845

Ser Ser Arg Leu Ala Gly Leu Gly Phe Arg lie lie Pro Leu Ala Cys 77 201038734 o 5 8 5 5 8 o 6 8 r h T a 1 A n _—_ G a 1 A a 5 17 A 8 r h T t e M s •1 H 0 r p r o e 7 s 8 s 1 H e h p a _—_ A y _—l G u 5 16 G 8 e o h 8 p 8Ser Ser Arg Leu Ala Gly Leu Gly Phe Arg lie lie Pro Leu Ala Cys 77 201038734 o 5 8 5 5 8 o 6 8 rh T a 1 A n ___ G a 1 A a 5 17 A 8 rh T te M s • 1 H 0 rproe 7 s 8 s 1 H ehpa _—_ A y _—l G u 5 16 G 8 eoh 8 p 8

Gin Ala Ala Leu Asp Ser Leu Lys lie Ser Thr Pro Thr Asn Gly Ala 885 890 895Gin Ala Ala Leu Asp Ser Leu Lys lie Ser Thr Pro Thr Asn Gly Ala 885 890 895

Arg Leu Tyr Asn Asn Val Ser Gly Lys Thr Cys Arg Ser Leu Gly Glu 900 905 910Arg Leu Tyr Asn Asn Val Ser Gly Lys Thr Cys Arg Ser Leu Gly Glu 900 905 910

Leu Arg Asp Cys Leu Gly Lys His Met Thr Ser Pro Val Leu Phe Gin 915 920 925Leu Arg Asp Cys Leu Gly Lys His Met Thr Ser Pro Val Leu Phe Gin 915 920 925

Ala Gin Val Glu Asn Met Tyr Ala Ala Gly Ala Arg lie Phe Val Glu 930 935 940 e 5 h4 p 9Ala Gin Val Glu Asn Met Tyr Ala Ala Gly Ala Arg lie Phe Val Glu 930 935 940 e 5 h4 p 9

Gly Pro Lys Gin Val Leu Ser Lys Leu Val Gly Glu lie Leu Ala 950 955 960Gly Pro Lys Gin Val Leu Ser Lys Leu Val Gly Glu lie Leu Ala 950 955 960

Asp Lys Ser Asp Phe Val Thr Val Ala Val Asn Ser Ser Ser Ser Lys 965 970 975Asp Lys Ser Asp Phe Val Thr Val Ala Val Asn Ser Ser Ser Ser Lys 965 970 975

Asp Ser Asp Val Gin Leu Arg Glu Ala Ala Ala Lys Leu Ala Val Leu 980 985 990Asp Ser Asp Val Gin Leu Arg Glu Ala Ala Ala Lys Leu Ala Val Leu 980 985 990

Gly Val Pro Leu Ala Asn Phe Asp Pro Trp Glu Leu Cys Asp Ala Arg 995 1000 1005Gly Val Pro Leu Ala Asn Phe Asp Pro Trp Glu Leu Cys Asp Ala Arg 995 1000 1005

Arg Leu Arg Glu Cys Pro Arg Ser Lys Thr Thr Leu Arg Leu Ser 1010 1015 1020Arg Leu Arg Glu Cys Pro Arg Ser Lys Thr Thr Leu Arg Leu Ser 1010 1015 1020

Ala Ala Thr Tyr Val Ser Asn Lys Thr Leu Ala Ala Arg Glu Lys 1025 1030 1035Ala Ala Thr Tyr Val Ser Asn Lys Thr Leu Ala Ala Arg Glu Lys 1025 1030 1035

Val Met Glu Asp Asn Cys Asp Phe Ser Ser Leu Phe Ala Ser Gly 1040 1045 1050Val Met Glu Asp Asn Cys Asp Phe Ser Ser Leu Phe Ala Ser Gly 1040 1045 1050

Pro Ala Ser Gin Glu Met Glu Arg Glu lie Ala Asn Leu Arg Ala 1055 1060 1065Pro Ala Ser Gin Glu Met Glu Arg Glu lie Ala Asn Leu Arg Ala 1055 1060 1065

Glu Leu Glu Ala Ala Gin Arg Gin Leu Asp Thr Ala Lys Thr Gin 1070 1075 1080Glu Leu Glu Ala Ala Gin Arg Gin Leu Asp Thr Ala Lys Thr Gin 1070 1075 1080

Leu Ala Arg Lys Gin Val Gin Asp Pro Thr Ala Asp Arg Gin Arg 1085 1090 1095Leu Ala Arg Lys Gin Val Gin Asp Pro Thr Ala Asp Arg Gin Arg 1085 1090 1095

Asp Met lie Ala Lys His Arg Ser Thr Leu Ala Ala Met Val Lys 1100 1105 1110Asp Met lie Ala Lys His Arg Ser Thr Leu Ala Ala Met Val Lys 1100 1105 1110

Glu Phe Glu Ala Leu Ala Ser Gly Ser Pro Cys Ala Val Pro Phe 1115 1120 1125Glu Phe Glu Ala Leu Ala Ser Gly Ser Pro Cys Ala Val Pro Phe 1115 1120 1125

Ala Pro Val Val Asp Thr Ala Val Glu Asp Val Pro Phe Ala Asp 1130 1135 1140 78 201038734Ala Pro Val Val Asp Thr Ala Val Glu Asp Val Pro Phe Ala Asp 1130 1135 1140 78 201038734

Lys Val Ser Thr Pro Pro Pro Gin Val Thr Ser Ala Pro lie Ala 1145 1150 1155Lys Val Ser Thr Pro Pro Pro Gin Val Thr Ser Ala Pro lie Ala 1145 1150 1155

Glu Leu Ala Arg Ala Glu Ala Val Val Met Glu Val Leu Ala Ala 1160 1165 1170Glu Leu Ala Arg Ala Glu Ala Val Val Met Glu Val Leu Ala Ala 1160 1165 1170

Lys Thr Gly Tyr Glu Val Asp Met lie Glu Ala Asp Met Leu Leu 1175 1180 1185Lys Thr Gly Tyr Glu Val Asp Met lie Glu Ala Asp Met Leu Leu 1175 1180 1185

Asp Ala Glu Leu Gly lie Asp Ser Val Lys Arg lie Glu lie Leu 1190 1195 1200Asp Ala Glu Leu Gly lie Asp Ser Val Lys Arg lie Glu lie Leu 1190 1195 1200

Ala Ala Val Gin Ala Gin Leu Gly Val Glu Ala Lys Asp Val Asp 1205 1210 1215Ala Ala Val Gin Ala Gin Leu Gly Val Glu Ala Lys Asp Val Asp 1205 1210 1215

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met 1220 1225 1230 ΟAla Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met 1220 1225 1230 Ο

Lys Ala Glu lie Gly Gly Gin Ala Thr Ser Ala Pro Ser Pro Met 1235 1240 1245Lys Ala Glu lie Gly Gly Gin Ala Thr Ser Ala Pro Ser Pro Met 1235 1240 1245

Ala Gin Pro Gin Ala Ser Ala Pro Ser Pro Ser Pro Thr Ala Ser 1250 1255 1260Ala Gin Pro Gin Ala Ser Ala Pro Ser Pro Ser Pro Thr Ala Ser 1250 1255 1260

Val Leu Pro Lys Pro Val Ala Leu Pro Ala Ser Val Asp Pro Ala 1265 1270 1275Val Leu Pro Lys Pro Val Ala Leu Pro Ala Ser Val Asp Pro Ala 1265 1270 1275

Lys Leu Ala Arg Ala Glu Ala Val Val Met Glu Val Leu Ala Ala 1280 1285 1290Lys Leu Ala Arg Ala Glu Ala Val Val Met Glu Val Leu Ala Ala 1280 1285 1290

Lys Thr Gly Tyr Glu Val Asp Met lie Glu Ala Asp Met Leu Leu 1295 1300 1305Lys Thr Gly Tyr Glu Val Asp Met lie Glu Ala Asp Met Leu Leu 1295 1300 1305

Asp Ala Glu Leu Gly lie Asp Ser Val Lys Arg lie Glu lie Leu 1310 1315 1320Asp Ala Glu Leu Gly lie Asp Ser Val Lys Arg lie Glu lie Leu 1310 1315 1320

Ala Ala Val Gin Ala Gin Leu Gly Val Glu Ala Lys Asp Val Asp 1325 1330 1335Ala Ala Val Gin Ala Gin Leu Gly Val Glu Ala Lys Asp Val Asp 1325 1330 1335

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met 1340 1345 1350Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met 1340 1345 1350

Lys Ala Glu lie Gly Gly Gin Ala Thr Ser Ala Pro Ala Ser Val 1355 1360 1365Lys Ala Glu lie Gly Gly Gin Ala Thr Ser Ala Pro Ala Ser Val 1355 1360 1365

Ala Gin Pro Gin Ala Ser Ala Pro Ser Pro Ser Ala Thr Thr Ala 1370 1375 1380Ala Gin Pro Gin Ala Ser Ala Pro Ser Pro Ser Ala Thr Thr Ala 1370 1375 1380

Ser Val Leu Pro Lys Pro Val Ala Ala Pro Thr Ser Ala Asp Pro 1385 1390 1395Ser Val Leu Pro Lys Pro Val Ala Ala Pro Thr Ser Ala Asp Pro 1385 1390 1395

Ala Lys Leu Ala Arg Ala Glu Ala Val Val Met Glu Val Leu Ala 1400 1405 1410Ala Lys Leu Ala Arg Ala Glu Ala Val Val Met Glu Val Leu Ala 1400 1405 1410

Ala Lys Thr Gly Tyr Glu Val Asp Met lie Glu Ala Asp Met Leu 1415 1420 1425 79 201038734Ala Lys Thr Gly Tyr Glu Val Asp Met lie Glu Ala Asp Met Leu 1415 1420 1425 79 201038734

Leu Asp Ala Glu Leu Gly lie Asp Ser Val Lys Arg lie Glu lie 1430 1435 1440Leu Asp Ala Glu Leu Gly lie Asp Ser Val Lys Arg lie Glu lie 1430 1435 1440

Leu Ala Ala Val Gin Ala Gin Leu Gly Val Glu Ala Lys Asp Val 1445 1450 1455Leu Ala Ala Val Gin Ala Gin Leu Gly Val Glu Ala Lys Asp Val 1445 1450 1455

Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Glu Ala 1460 1465 1470Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val Val Glu Ala 1460 1465 1470

Met Lys Ala Glu lie Gly Gly Gin Ala Thr Ser Ala Pro Ala Ser 1475 1480 1485Met Lys Ala Glu lie Gly Gly Gin Ala Thr Ser Ala Pro Ala Ser 1475 1480 1485

Val Ala Gin Pro Gin lie Ser Val Ser Pro Thr Pro Leu Ala Ala 1490 1495 1500Val Ala Gin Pro Gin lie Ser Val Ser Pro Thr Pro Leu Ala Ala 1490 1495 1500

Ser Pro Ser Ala Asp Pro Ala Lys Leu Ala Arg Ala Glu Ala Val 1505 1510 1515Ser Pro Ser Ala Asp Pro Ala Lys Leu Ala Arg Ala Glu Ala Val 1505 1510 1515

Val Met Glu Val Leu Ala Ala Lys Thr Gly Tyr Glu Val Asp Met 1520 1525 1530 lie Glu Ala Asp Met Leu Leu Asp Ala Glu Leu Gly lie Asp Ser 1535 1540 1545Val Met Glu Val Leu Ala Ala Lys Thr Gly Tyr Glu Val Asp Met 1520 1525 1530 lie Glu Ala Asp Met Leu Leu Asp Ala Glu Leu Gly lie Asp Ser 1535 1540 1545

Val Lys Arg lie Glu lie Leu Ala Ala Val Gin Ala Gin Leu Gly 1550 1555 1560Val Lys Arg lie Glu lie Leu Ala Ala Val Gin Ala Gin Leu Gly 1550 1555 1560

Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val 1565 1570 1575Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val 1565 1570 1575

Gly Glu Val Val Asp Ala Met Lys Ala Glu lie Gly Gly Gin Ala 1580 1585 1590Gly Glu Val Val Asp Ala Met Lys Ala Glu lie Gly Gly Gin Ala 1580 1585 1590

Thr Ser Ala Pro Ala Ser Val Ala Gin Pro Gin Ala Ser Ala Pro 1595 1600 1605Thr Ser Ala Pro Ala Ser Val Ala Gin Pro Gin Ala Ser Ala Pro 1595 1600 1605

Ser Pro Ser Ala Thr Ala Ser Val Leu Pro Lys Pro Val Ala Ala 1610 1615 1620Ser Pro Ser Ala Thr Ala Ser Val Leu Pro Lys Pro Val Ala Ala 1610 1615 1620

Pro Thr Ser Ala Asp Pro Ala Lys Leu Ala Arg Ala Glu Ala Val 1625 1630 1635Pro Thr Ser Ala Asp Pro Ala Lys Leu Ala Arg Ala Glu Ala Val 1625 1630 1635

Val Met Glu Val Leu Ala Ala Lys Thr Gly Tyr Glu Val Asp Met 1640 1645 1650 lie Glu Ala Asp Met Leu Leu Asp Ala Glu Leu Gly lie Asp Ser 1655 1660 1665Val Met Glu Val Leu Ala Ala Lys Thr Gly Tyr Glu Val Asp Met 1640 1645 1650 lie Glu Ala Asp Met Leu Leu Asp Ala Glu Leu Gly lie Asp Ser 1655 1660 1665

Val Lys Arg He Glu He Leu Ala Ala Val Gin Ala Gin Leu Gly 1670 1675 1680Val Lys Arg He Glu He Leu Ala Ala Val Gin Ala Gin Leu Gly 1670 1675 1680

Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val 1685 1690 1695Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val 1685 1690 1695

Gly Glu〇 Val Val Glu Ala Me^ Lys Ala Glu He Gly〇 Gly Gin Ala 80 201038734Gly Glu〇 Val Val Glu Ala Me^ Lys Ala Glu He Gly〇 Gly Gin Ala 80 201038734

Thr Ser Ala Pro Ala Ser Met Ala Gin Pro Gin lie Ser Val Ser 1715 1720 1725Thr Ser Ala Pro Ala Ser Met Ala Gin Pro Gin lie Ser Val Ser 1715 1720 1725

Pro Thr Pro Leu Ala Ala Ser Pro Ser Ala Asp Pro Ala Lys Leu 1730 1735 1740Pro Thr Pro Leu Ala Ala Ser Pro Ser Ala Asp Pro Ala Lys Leu 1730 1735 1740

Ala Arg Ala Glu Ala Val Val Met Glu Val Leu Ala Ala Lys Thr 1745 1750 1755Ala Arg Ala Glu Ala Val Val Met Glu Val Leu Ala Ala Lys Thr 1745 1750 1755

Gly Tyr Glu Val Asp Met lie Glu Ala Asp Met Leu Leu Asp Ala 1760 1765 1770Gly Tyr Glu Val Asp Met lie Glu Ala Asp Met Leu Leu Asp Ala 1760 1765 1770

Glu Leu Gly lie Asp Ser Val Lys Arg lie Glu lie Leu Ala Ala 1775 1780 1785Glu Leu Gly lie Asp Ser Val Lys Arg lie Glu lie Leu Ala Ala 1775 1780 1785

ΟΟ

Val Gin Ala Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu 1790 1795 1800Val Gin Ala Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu 1790 1795 1800

Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala 1805 1810 1815Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala 1805 1810 1815

Glu lie Gly Gly Gin Ala Thr Ser Ala Pro Ala Ser Val Ala Gin 1820 1825 1830Glu lie Gly Gly Gin Ala Thr Ser Ala Pro Ala Ser Val Ala Gin 1820 1825 1830

Pro Gin Ala Ser Ala Pro Ser Pro Ser Ala Thr Ala Ser Ala Pro 1835 1840 1845Pro Gin Ala Ser Ala Pro Ser Pro Ser Ala Thr Ala Ser Ala Pro 1835 1840 1845

Val Thr Pro Leu Ala Ala Pro Ala Ser Val Asp Pro Ala Lys Leu 1850 1855 1860Val Thr Pro Leu Ala Ala Pro Ala Ser Val Asp Pro Ala Lys Leu 1850 1855 1860

Ala Arg Ala Glu Ala Val Val Met Glu Val Leu Ala Ala Lys Thr 1865 1870 1875Ala Arg Ala Glu Ala Val Val Met Glu Val Leu Ala Ala Lys Thr 1865 1870 1875

Gly Tyr Glu Val Asp Met lie Glu Ala Asp Met Leu Leu Asp Ala 1880 1885 1890Gly Tyr Glu Val Asp Met lie Glu Ala Asp Met Leu Leu Asp Ala 1880 1885 1890

Glu Leu Gly lie Asp Ser Val Lys Arg lie Glu lie Leu Ala Ala 1895 1900 1905Glu Leu Gly lie Asp Ser Val Lys Arg lie Glu lie Leu Ala Ala 1895 1900 1905

Val Gin Ala Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu 1910 1915 1920Val Gin Ala Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu 1910 1915 1920

Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala 1925 1930 1935Ser Arg Thr Arg Thr Val Gly Glu Val Val Asp Ala Met Lys Ala 1925 1930 1935

Glu lie Gly Gly Gin Ala Thr Ser Ala Pro Ala Ser Val Ala Gin 1940 1945 1950Glu lie Gly Gly Gin Ala Thr Ser Ala Pro Ala Ser Val Ala Gin 1940 1945 1950

Pro Gin Ala Ser Ala Pro Ser Pro Ser Ala Thr Ala Ser Val Leu 1955 1960 1965Pro Gin Ala Ser Ala Pro Ser Pro Ser Ala Thr Ala Ser Val Leu 1955 1960 1965

Pro Lys Pro Val Ala Ser Pro Ala Ser Val Asp Pro Ala Lys Leu 1970 1975 1980Pro Lys Pro Val Ala Ser Pro Ala Ser Val Asp Pro Ala Lys Leu 1970 1975 1980

Ala Arg Ala Glu Ala Val Val Met Glu Val Leu Ala Ala Lys Thr 81 201038734 1985 1990 1995Ala Arg Ala Glu Ala Val Val Met Glu Val Leu Ala Ala Lys Thr 81 201038734 1985 1990 1995

Gly Tyr Glu Val Asp Met lie Asp Ala Asp Met Leu Leu Asp Ala 2000 2005 2010Gly Tyr Glu Val Asp Met lie Asp Ala Asp Met Leu Leu Asp Ala 2000 2005 2010

Glu Leu Gly lie Asp Ser Val Lys Arg lie Glu lie Leu Ala Ala 2015 2020 2025Glu Leu Gly lie Asp Ser Val Lys Arg lie Glu lie Leu Ala Ala 2015 2020 2025

Val Gin Ala Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu 2030 2035 2040Val Gin Ala Gin Leu Gly Val Glu Ala Lys Asp Val Asp Ala Leu 2030 2035 2040

Ser Arg Thr Arg Thr Val Gly Glu Val Val Glu Ala Met Lys Ala 2045 2050 2055Ser Arg Thr Arg Thr Val Gly Glu Val Val Glu Ala Met Lys Ala 2045 2050 2055

Glu lie Gly Ala Ala Gly Pro Asn Asp Ala Gin Ala Ala Ser Gly 2060 2065 2070Glu lie Gly Ala Ala Gly Pro Asn Asp Ala Gin Ala Ala Ser Gly 2060 2065 2070

His Leu Phe Gly Thr Gly Cys Glu Asp Leu Ser Leu Cys Ser Ala 2075 2080 2085His Leu Phe Gly Thr Gly Cys Glu Asp Leu Ser Leu Cys Ser Ala 2075 2080 2085

Ser Val Val Glu lie Ala Arg Cys Ser Glu Leu Ala Leu Glu Arg 2090 2095 2100Ser Val Val Glu lie Ala Arg Cys Ser Glu Leu Ala Leu Glu Arg 2090 2095 2100

Pro Met Asp Arg Pro lie Leu lie Val Ser Asp Gly Ser Ala Leu 2105 2110 2115Pro Met Asp Arg Pro lie Leu lie Val Ser Asp Gly Ser Ala Leu 2105 2110 2115

Pro Ala Ala Leu Ala Ser Arg Leu Gly Ser Cys Ala Val lie Leu 2120 2125 2130Pro Ala Ala Leu Ala Ser Arg Leu Gly Ser Cys Ala Val lie Leu 2120 2125 2130

Thr Thr Ala Gly Glu Thr Asp Gin Ser Val Arg Ser Thr Lys His 2135 2140 2145Thr Thr Ala Gly Glu Thr Asp Gin Ser Val Arg Ser Thr Lys His 2135 2140 2145

Val Asp Met Glu Gly Trp Gly Glu Ala Asp Leu Val Arg Ala Leu 2150 2155 2160Val Asp Met Glu Gly Trp Gly Glu Ala Asp Leu Val Arg Ala Leu 2150 2155 2160

Glu Ala Val Glu Ser Arg Phe Gly Val Pro Gly Gly Val Val Val 2165 2170 2175Glu Ala Val Glu Ser Arg Phe Gly Val Pro Gly Gly Val Val Val 2165 2170 2175

Leu Glu Arg Ala Ser Glu Thr Ala Arg Asp Gin Leu Gly Phe Ala 2180 2185 2190Leu Glu Arg Ala Ser Glu Thr Ala Arg Asp Gin Leu Gly Phe Ala 2180 2185 2190

Leu Leu Leu Ala Lys His Ser Ser Lys Ala Leu Asn Gin Gin lie 2195 2200 2205Leu Leu Leu Ala Lys His Ser Ser Lys Ala Leu Asn Gin Gin lie 2195 2200 2205

Pro Gly Gly Arg Ala Cys Phe Val Gly Val Ser Arg lie Asp Gly 2210 2215 2220Pro Gly Gly Arg Ala Cys Phe Val Gly Val Ser Arg lie Asp Gly 2210 2215 2220

Lys Leu Gly Leu Ser Gly Ala Cys Ala Lys Gly Lys Gly Trp Ala 2225 2230 2235Lys Leu Gly Leu Ser Gly Ala Cys Ala Lys Gly Lys Gly Trp Ala 2225 2230 2235

Glu Ala. Al3. Glu 11 θ A1 a. Gin Gin Gly Als Vsl Ala. Gly Leu Cys 2240 2245 2250Glu Ala. Al3. Glu 11 θ A1 a. Gin Gin Gly Als Vsl Ala. Gly Leu Cys 2240 2245 2250

Lys Thr Leu Asp Leu Glu Trp Pro His Val Phe Ala Arg Ser lie 2255 2260 2265 82 201038734Lys Thr Leu Asp Leu Glu Trp Pro His Val Phe Ala Arg Ser lie 2255 2260 2265 82 201038734

Asp lie Glu Leu Gly Ala 2270 Phe Glu Glu Leu Ser Cys 2285 Tyr Thr Lys Asp Gly Lys 2300 Gly Leu Gly Lys Pro Lys 2315 Leu Val Ser Gly Gly Ala 2330 Glu Leu Ala Lys Ser lie 2345 Glu Ala Asn lie Gly Thr 2375 Phe Ala Ala Gly Arg Gly 2390 Ala Leu Val Gly Ser Val 2405 Leu Glu Ser lie Arg Ala 2420 Cys Asp Val Ser Cys Ala 2435 Leu Glu Arg Arg Val Gly Gly Val Leu Arg Asp Lys 2465 Phe Glu Val Val Tyr Gly 2480 Leu Gin Ala Val Asp Arg 2495 Ser Ser Leu Ala Gly Phe 2510 Ala Met Ala Asn Glu Ala 2525 Thr Ala Met Pro Gly Leu 2540 Asn 2275 Glu Glu Thr Ala Ala 2280 Gin Ala lie Pro 2290 Asp Leu Thr Val Arg 2295 Glu Ala Gly Arg 2305 Trp Thr Thr Glu Ala 2310 Arg Pro Val Gin 2320 Ala Leu Arg Ser Ser 2325 Asp Val Phe Arg 2335 Gly lie Thr Pro Val 2340 Cys Val Arg Ser 2350 Gly Gly Thr Phe Val 2355 Leu Leu Gly Asp 2365 Pro Ala Trp Ala Cys 2370 Gly Val Glu Ala 2380 Ala Met Ala His Leu 2385 Lys Ala Glu Pro 2395 Lys Pro Thr Pro Lys 2400 Ala His Lys Leu 2410 Gly Ala Arg Glu Val 2415 Leu Gly Ser Gin 2425 Gly Ala Arg Ala Glu 2430 Tyr Val Ser Glu 2440 Arg Val Lys Ala Val 2445 Val Asp Asp Ala 2455 Val Thr Gly Val Val 2460 His Ala Ser Ser 2470 Val Glu Arg Leu Glu 2475 Leu Ala Asp Thr 2485 Lys Val Asp Gly Leu 2490 Leu Asn Leu Pro 2500 Lys Leu Arg His Leu 2505 Val Leu Phe His 2515 Gly Asn Thr Gly Gin 2520 Ala Val Tyr Leu 2530 Asn Lys Met Ala Phe 2535 His Leu Glu Ser 2545 Val Lys Thr lie Gly 2550 Phe Gly Pro 83 201038734Asp lie Glu Leu Gly Ala 2270 Phe Glu Glu Leu Ser Cys 2285 Tyr Thr Lys Asp Gly Lys 2300 Gly Leu Gly Lys Pro Lys 2315 Leu Val Ser Gly Gly Ala 2330 Glu Leu Ala Lys Ser lie 2345 Glu Ala Asn lie Gly Thr 2375 Phe Ala Ala Gly Arg Gly 2390 Ala Leu Val Gly Ser Val 2405 Leu Glu Ser lie Arg Ala 2420 Cys Asp Val Ser Cys Ala 2435 Leu Glu Arg Arg Val Gly Gly Val Leu Arg Asp Lys 2465 Phe Glu Val Val Tyr Gly 2480 Leu Gin Ala Val Asp Arg 2495 Ser Ser Leu Ala Gly Phe 2510 Ala Met Ala Asn Glu Ala 2525 Thr Ala Met Pro Gly Leu 2540 Asn 2275 Glu Glu Thr Ala Ala 2280 Gin Ala lie Pro 2290 Asp Leu Thr Val Arg 2295 Glu Ala Gly Arg 2305 Trp Thr Thr Glu Ala 2310 Arg Pro Val Gin 2320 Ala Leu Arg Ser Ser 2325 Asp Val Phe Arg 2335 Gly lie Thr Pro Val 2340 Cys Val Arg Ser 2350 Gly Gly Thr Phe Val 2355 Leu Leu Gly Asp 2365 Pro Ala Trp Ala Cys 2370 Gly Val Glu Ala 2380 Ala Met Ala His Leu 2385 Lys Ala Glu Pro 2395 Lys Pro Thr Pro Lys 2400 Ala His Lys Leu 2410 Gly Ala Arg Glu Val 2415 Leu Gly Ser Gin 2425 Gl y Ala Arg Ala Glu 2430 Tyr Val Ser Glu 2440 Arg Val Lys Ala Val 2445 Val Asp Asp Ala 2455 Val Thr Gly Val Val 2460 His Ala Ser Ser 2470 Val Glu Arg Leu Glu 2475 Leu Ala Asp Thr 2485 Lys Val Asp Gly Leu 2490 Leu Asn Leu Pro 2500 Lys Leu Arg His Leu 2505 Val Leu Phe His 2515 Gly Asn Thr Gly Gin 2520 Ala Val Tyr Leu 2530 Asn Lys Met Ala Phe 2535 His Leu Glu Ser 2545 Val Lys Thr lie Gly 2550 Phe Gly Pro 83 201038734

Trp Asp Gly Gly 2555 Met Val Asn Asp Ala Leu Lys Ala His Phe Ala 2560 2565 Ser Met Gly Val 2570 Gin lie lie Pro Leu Asp Gly Gly Ala Glu Thr 2575 2580 Val Ser Arg lie 2585 lie Gly Ala Cys Ser Pro Thr Gin Val Leu Val 2590 2595 Gly Asn Trp Gly 2600 Leu Pro Pro Val Val Pro Asn Ala Ser Val His 2605 2610 Lys lie Thr Val 2615 Arg Leu Gly Gly Glu Ser Ala Asn Pro Phe Leu 2620 2625 Ser Ser His Thr 2630 工le Gin Gly Arg Lys Val Leu Pro Met Thr Val 2635 2640 Ala Leu Gly Leu 2645 Leu Ala Glu Ala Ala Arg Gly Leu Tyr Val Gly 2650 2655 His Gin Val Val 2660 Gly lie Glu Asp Ala Gin Val Phe Gin Gly Val 2665 2670 Val Leu Asp Lys 2675 Gly Ala Thr Cys Glu Val Gin Leu Arg Arg Glu 2680 2685 Ser Ser Thr Ala 2690 Ser Pro Ser Glu Val Val Leu Ser Ala Ser Leu 2695 2700 Asn Val Phe Ala 2705 Ala Gly Lys Val Val Pro Ala Tyr Arg Ala His 2710 2715Trp Asp Gly Gly 2555 Met Val Asn Asp Ala Leu Lys Ala His Phe Ala 2560 2565 Ser Met Gly Val 2570 Gin lie lie Pro Leu Asp Gly Gly Ala Glu Thr 2575 2580 Val Ser Arg lie 2585 lie Gly Ala Cys Ser Pro Thr Gin Val Leu Val 2590 2595 Gly Asn Trp Gly 2600 Leu Pro Pro Val Val Pro Asn Ala Ser Val His 2605 2610 Lys lie Thr Val 2615 Arg Leu Gly Gly Glu Ser Ala Asn Pro Phe Leu 2620 2625 Ser Ser His Thr 2630 work le Gin Gly Arg Lys Val Leu Pro Met Thr Val 2635 2640 Ala Leu Gly Leu 2645 Leu Ala Glu Ala Ala Arg Gly Leu Tyr Val Gly 2650 2655 His Gin Val Val 2660 Gly lie Glu Asp Ala Gin Val Phe Gin Gly Val 2665 2670 Val Leu Asp Lys 2675 Gly Ala Thr Cys Glu Val Gin Leu Arg Arg Glu 2680 2685 Ser Ser Thr Ala 2690 Ser Pro Ser Glu Val Val Leu Ser Ala Ser Leu 2695 2700 Asn Val Phe Ala 2705 Ala Gly Lys Val Val Pro Ala Tyr Arg Ala His 2710 2715

Val Val Leu Gly Ala Ser Gly Pro Arg Thr Gly Gly Val Gin Leu 2720 2725 2730Val Val Leu Gly Ala Ser Gly Pro Arg Thr Gly Gly Val Gin Leu 2720 2725 2730

Glu Leu Lys Asp 2735 Leu Gly Val Asp Ala Asp Pro Ala Cys Ser Val 2740 2745 Gly Lys Gly Ala 2750 Leu Tyr Asp Gly Arg Thr Leu Phe His Gly Pro 2755 2760 Ala Phe Gin Tyr 2765 Met Asp Glu Val Leu Arg Cys Ser Pro Ala Glu 2770 2775 Leu Ala Val Arg 2780 Cys Arg Val Val Pro Ser Ala Ala Gin Asp Arg 2785 2790 Gly Gin Phe Val 2795 Ser Arg Gly Val Leu Tyr Asp Pro Phe Leu Asn 2800 2805 Asp Thr Val Phe 2810 Gin Ala Leu Leu Val Trp Ala Arg Leu Val Arg 2815 2820 Asp Ser Ala Ser 2825 Leu Pro Ser Asn Val Glu Arg lie Ser Phe His 2830 2835 84 201038734Glu Leu Lys Asp 2735 Leu Gly Val Asp Ala Asp Pro Ala Cys Ser Val 2740 2745 Gly Lys Gly Ala 2750 Leu Tyr Asp Gly Arg Thr Leu Phe His Gly Pro 2755 2760 Ala Phe Gin Tyr 2765 Met Asp Glu Val Leu Arg Cys Ser Pro Ala Glu 2770 2775 Leu Ala Val Arg 2780 Cys Arg Val Val Pro Ser Ala Ala Gin Asp Arg 2785 2790 Gly Gin Phe Val 2795 Ser Arg Gly Val Leu Tyr Asp Pro Phe Leu Asn 2800 2805 Asp Thr Val Phe 2810 Gin Ala Leu Leu Val Trp Ala Arg Leu Val Arg 2815 2820 Asp Ser Ala Ser 2825 Leu Pro Ser Asn Val Glu Arg lie Ser Phe His 2830 2835 84 201038734

Gly Gin Pro Pro Ser Glu Gly Glu Val Phe Tyr Thr Thr Leu Lys 2840 2845 2850Gly Gin Pro Pro Ser Glu Gly Glu Val Phe Tyr Thr Thr Leu Lys 2840 2845 2850

Leu Asp Ser Ala Ala Ser Gly Pro Leu Asp Pro lie Ala Lys Ala 2855 2860 2865Leu Asp Ser Ala Ala Ser Gly Pro Leu Asp Pro lie Ala Lys Ala 2855 2860 2865

Gin Phe Phe Leu His Arg Ala Cys Gly Ala Val Phe Ala Ser Gly 2870 2875 2880Gin Phe Phe Leu His Arg Ala Cys Gly Ala Val Phe Ala Ser Gly 2870 2875 2880

Arg Ala Ser Val Val Leu Asn Lys Ala Leu Ser Phe 2885 2890 2895 <210> 56 <211> 2811 <212> PRT <213〉 破囊壺菌物種Arg Ala Ser Val Val Leu Asn Lys Ala Leu Ser Phe 2885 2890 2895 <210> 56 <211> 2811 <212> PRT <213> Thraustochytrium Species

<400> 56<400> 56

Met Lys Asp Met Glu Asp Arg Arg Val Ala lie Val Gly Met Ser Ala 15 10 15Met Lys Asp Met Glu Asp Arg Arg Val Ala lie Val Gly Met Ser Ala 15 10 15

His Leu Pro Cys Gly Thr Asp Val Lys Glu Ser Trp Gin Ala lie Arg 20 25 30His Leu Pro Cys Gly Thr Asp Val Lys Glu Ser Trp Gin Ala lie Arg 20 25 30

Asp Gly lie Asp Cys Leu Ser Asp Leu Pro Ala Asp Arg Leu Asp Val 35 40 45Asp Gly lie Asp Cys Leu Ser Asp Leu Pro Ala Asp Arg Leu Asp Val 35 40 45

Thr Ala Tyr Tyr Asn Pro Asn Lys Ala Thr Lys Asp Lys lie Tyr Cys 50 55 60Thr Ala Tyr Tyr Asn Pro Asn Lys Ala Thr Lys Asp Lys lie Tyr Cys 50 55 60

Lys Arg Gly Gly Phe lie Pro Asn Tyr Asp Phe Asp Pro Arg Glu Phe 65 70 75 80Lys Arg Gly Gly Phe lie Pro Asn Tyr Asp Phe Asp Pro Arg Glu Phe 65 70 75 80

Gly Leu Asn Met Phe Gin Met Glu Asp Ser Asp Ala Asn Gin Thr Leu 85 90 95Gly Leu Asn Met Phe Gin Met Glu Asp Ser Asp Ala Asn Gin Thr Leu 85 90 95

Thr Leu Leu Lys Val Lys Gin Ala Leu Glu Asp Ala Ser lie Glu Pro 100 105 110Thr Leu Leu Lys Val Lys Gin Ala Leu Glu Asp Ala Ser lie Glu Pro 100 105 110

Phe Thr Lys Glu Lys Lys Asn lie Gly Cys Val Leu Gly lie Gly Gly 115 120 125Phe Thr Lys Glu Lys Lys Asn lie Gly Cys Val Leu Gly lie Gly Gly 115 120 125

Gly Gin Lys Ala Ser His Glu Phe Tyr Ser Arg Leu Asn Tyr Val Val 130 135 140Gly Gin Lys Ala Ser His Glu Phe Tyr Ser Arg Leu Asn Tyr Val Val 130 135 140

Val Glu Lys Val Leu Arg Lys Met Gly Leu Pro Asp Ala Asp Val Glu 145 150 155 160Val Glu Lys Val Leu Arg Lys Met Gly Leu Pro Asp Ala Asp Val Glu 145 150 155 160

Glu Ala Val Glu Lys Tyr Lys Ala Asn Phe Pro Glu Trp Arg Leu Asp 165 170 175Glu Ala Val Glu Lys Tyr Lys Ala Asn Phe Pro Glu Trp Arg Leu Asp 165 170 175

Ser Phe Pro Gly Phe Leu Gly Asn Val Thr Ala Gly Arg Cys Ser Asn 180 185 190Ser Phe Pro Gly Phe Leu Gly Asn Val Thr Ala Gly Arg Cys Ser Asn 180 185 190

Thr Phe Asn Met Glu Gly Met Asn Cys Val Val Asp Ala Ala Cys Ala 85 201038734 195 200 205Thr Phe Asn Met Glu Gly Met Asn Cys Val Val Asp Ala Ala Cys Ala 85 201038734 195 200 205

Ser Ser Leu lie Ala 工le Lys Val Ala Val Glu Glu Leu Leu Phe Gly 210 215 220Ser Ser Leu lie Ala l Lys Val Ala Val Glu Glu Leu Leu Phe Gly 210 215 220

Asp Cys Asp Thr Met lie Ala Gly Ala Thr Cys Thr Asp Asn Ser Leu 225 230 235 240Asp Cys Asp Thr Met lie Ala Gly Ala Thr Cys Thr Asp Asn Ser Leu 225 230 235 240

Gly Met Tyr Met Ala Phe Ser Lys Thr Pro Val Phe Ser Thr Asp Pro 245 250 255Gly Met Tyr Met Ala Phe Ser Lys Thr Pro Val Phe Ser Thr Asp Pro 245 250 255

Ser Val Arg Ala Tyr Asp Glu Lys Thr Lys Gly Met Leu lie Gly Glu 260 265 270Ser Val Arg Ala Tyr Asp Glu Lys Thr Lys Gly Met Leu lie Gly Glu 260 265 270

Gly Ser Ala Met Phe Val Leu Lys Arg Tyr Ala Asp Ala Val Arg Asp 275 280 285Gly Ser Ala Met Phe Val Leu Lys Arg Tyr Ala Asp Ala Val Arg Asp 275 280 285

Gly Asp Thr lie His Ala Val Leu Arg Ser Cys Ser Ser Ser Ser Asp 290 295 300Gly Asp Thr lie His Ala Val Leu Arg Ser Cys Ser Ser Ser Ser Asp 290 295 300

Gly Lys Ala Ala Gly lie Tyr Thr Pro Thr lie Ser Gly Gin Glu Glu 305 310 315 320Gly Lys Ala Ala Gly lie Tyr Thr Pro Thr lie Ser Gly Gin Glu Glu 305 310 315 320

Ala Leu Arg Arg Ala Tyr Ala Arg Ala Gly Val Cys Pro Ser Thr lie 325 330 335Ala Leu Arg Arg Ala Tyr Ala Arg Ala Gly Val Cys Pro Ser Thr lie 325 330 335

Gly Leu Val Glu Gly His Gly Thr Gly Thr Pro Val Gly Asp Arg lie 340 345 350Gly Leu Val Glu Gly His Gly Thr Gly Thr Pro Val Gly Asp Arg lie 340 345 350

Glu Leu Thr Ala Leu Arg Asn Leu Phe Asp Lys Ala Phe Gly Ser Lys 355 360 365Glu Leu Thr Ala Leu Arg Asn Leu Phe Asp Lys Ala Phe Gly Ser Lys 355 360 365

Lys Glu Gin lie Ala Val Gly Ser 工le Lys Ser Gin lie Gly His Leu 370 375 380Lys Glu Gin lie Ala Val Gly Ser Le Lys Ser Gin lie Gly His Leu 370 375 380

Lys Ser Val Ala Gly Phe Ala Gly Leu Val Lys Ala Val Leu Ala Leu 385 390 395 400Lys Ser Val Ala Gly Phe Ala Gly Leu Val Lys Ala Val Leu Ala Leu 385 390 395 400

Lys His Lys Thr Leu Pro Gly Ser lie Asn Val Asp Gin Pro Pro Leu 405 410 415Lys His Lys Thr Leu Pro Gly Ser lie Asn Val Asp Gin Pro Pro Leu 405 410 415

Leu Tyr Asp Gly Thr Gin lie Gin Asp Ser Ser Leu Tyr lie Asn Lys 420 425 430Leu Tyr Asp Gly Thr Gin lie Gin Asp Ser Ser Leu Tyr lie Asn Lys 420 425 430

Thr Asn Arg Pro Trp Phe Thr Gin Asn Lys Leu Pro Ara Ara Ala Gly 435 440 445Thr Asn Arg Pro Trp Phe Thr Gin Asn Lys Leu Pro Ara Ara Ala Gly 435 440 445

Val Ser Ser Phe Gly Phe Gly Gly Ala Asn Tyr His Ala Val Leu Glu 450 455 460Val Ser Ser Phe Gly Phe Gly Gly Ala Asn Tyr His Ala Val Leu Glu 450 455 460

Glu Phe Glu Pro Glu His Glu Lys Pro Tyr Arg Leu Asn Thr Val Gly 465 470 475 480Glu Phe Glu Pro Glu His Glu Lys Pro Tyr Arg Leu Asn Thr Val Gly 465 470 475 480

His Pro Val Leu Leu Tyr Ala Pro Ser Val Glu Ala Leu Lys Val Leu 485 490 495 86 201038734His Pro Val Leu Leu Tyr Ala Pro Ser Val Glu Ala Leu Lys Val Leu 485 490 495 86 201038734

Cys Asn Asp Gin Leu Ala Glu Leu 500Cys Asn Asp Gin Leu Ala Glu Leu 500

Thr lie Ala Leu Glu Glu Ala 505 510Thr lie Ala Leu Glu Glu Ala 505 510

Thr His Lys Asn Val Asp Lys Val 515 520Thr His Lys Asn Val Asp Lys Val 515 520

Cys Gly Tyr Lys Phe 工le Asp 525Cys Gly Tyr Lys Phe work le Asp 525

Phe Gin Leu Gin Gly Ser Cys Pro 530 535Phe Gin Leu Gin Gly Ser Cys Pro 530 535

Pro Glu Asn Pro Arg Val Gly 540Pro Glu Asn Pro Arg Val Gly 540

Leu Ala Thr Leu Pro Thr Ser Asn 545 550 lie lie Val Ala Leu Lys Ala 555Leu Ala Thr Leu Pro Thr Ser Asn 545 550 lie lie Val Ala Leu Lys Ala 555

Leu Ala Gin Leu Asp Ala Lys Pro 565Leu Ala Gin Leu Asp Ala Lys Pro 565

Asp Ala Lys Lys Trp Asp Leu 570 575Asp Ala Lys Lys Trp Asp Leu 570 575

His Lys Lys Ala Phe Gly Ala Thr 580His Lys Lys Ala Phe Gly Ala Thr 580

Phe Ala Ser Ser Ser Val Lys 585 590 ΟPhe Ala Ser Ser Ser Val Lys 585 590 Ο

Ser Val Ala Ala Leu Phe Ala Gly 595 600Ser Val Ala Ala Leu Phe Ala Gly 595 600

Gin Gly Thr Gin Tyr Leu Asn 605Gin Gly Thr Gin Tyr Leu Asn 605

Phe Ser Asp Val Ala Met Asn Trp 610 615Phe Ser Asp Val Ala Met Asn Trp 610 615

Pro Pro Phe Arg Asp Ser lie 620Pro Pro Phe Arg Asp Ser lie 620

Ala Met Glu Glu Ala Gin Thr Glu 625 630Ala Met Glu Glu Ala Gin Thr Glu 625 630

Val Phe Glu Gly Gin Val Glu 635 lie Ser Lys Val Leu Phe Pro Arg 645Val Phe Glu Gly Gin Val Glu 635 lie Ser Lys Val Leu Phe Pro Arg 645

Glu Arg Tyr Ala Ser Glu Ser 650 655Glu Arg Tyr Ala Ser Glu Ser 650 655

Gin Gly Asn Glu Leu Leu Cys Leu 660Gin Gly Asn Glu Leu Leu Cys Leu 660

Thr Glu Tyr Ser Gin Pro Thr 665 670 lie Ala Ala Ala Val Gly Ala Phe 675 680Thr Glu Tyr Ser Gin Pro Thr 665 670 lie Ala Ala Ala Val Gly Ala Phe 675 680

Asp lie Phe Lys Ala Ala Gly 685Asp lie Phe Lys Ala Ala Gly 685

Lys Pro Asp Met Val Gly Gly His 690 695Lys Pro Asp Met Val Gly Gly His 690 695

Ser Leu Gly Glu Phe Ala Ala 700Ser Leu Gly Glu Phe Ala Ala 700

Tyr Ala Ala Gly Ser lie Ser Arg 705 710Tyr Ala Ala Gly Ser lie Ser Arg 705 710

Asp Asp Leu Tyr Lys Leu Val 715Asp Asp Leu Tyr Lys Leu Val 715

Lys Arg Ala Lys Ala Met Ala Asn 725Lys Arg Ala Lys Ala Met Ala Asn 725

Ala Ser Asp Gly Ala Met Ala 730 735Ala Ser Asp Gly Ala Met Ala 730 735

Val 工le Gly Pro Asp Ala Arg Leu *7 40Val work le Gly Pro Asp Ala Arg Leu *7 40

Val Thr Pro Gin Asn Ser Asp 745 750Val Thr Pro Gin Asn Ser Asp 745 750

Tyr Val Ala Asn Phe Asn Ser Ala 755 760Tyr Val Ala Asn Phe Asn Ser Ala 755 760

Thr Gin Val Val lie Ser Gly 765Thr Gin Val Val lie Ser Gly 765

Val Gin Gly Val Lys Glu Glu Ser 770 775Val Gin Gly Val Lys Glu Glu Ser 770 775

Lys Leu Leu lie Ser Lys Gly 780Lys Leu Leu lie Ser Lys Gly 780

Arg Val Leu Pro Leu Lys Cys Gin 785 790Arg Val Leu Pro Leu Lys Cys Gin 785 790

Gly Ala Phe His Ser Pro Leu 795Gly Ala Phe His Ser Pro Leu 795

Lys Glu Phe lie 560 Pro Gly Met Val Pro 640 Glu Thr Phe Leu Cys 720 Ala Val Thr Phe Met 800 87 201038734Lys Glu Phe lie 560 Pro Gly Met Val Pro 640 Glu Thr Phe Leu Cys 720 Ala Val Thr Phe Met 800 87 201038734

Gly Pro Ser Glu Asp Ser Phe Lys Ser Leu Val Glu Thr Cys Thr lie 805 810 815Gly Pro Ser Glu Asp Ser Phe Lys Ser Leu Val Glu Thr Cys Thr lie 805 810 815

Ser Pro Pro Lys Asn Val Lys Phe Phe Cys Asn Val Ser Gly Lys Glu 820 825 830Ser Pro Pro Lys Asn Val Lys Phe Phe Cys Asn Val Ser Gly Lys Glu 820 825 830

Ser Pro Asn Pro Lys Gin Thr Leu Lys Ser His Met Thr Ser Ser Val 835 840 845Ser Pro Asn Pro Lys Gin Thr Leu Lys Ser His Met Thr Ser Ser Val 835 840 845

Gin Phe Glu Glu Gin lie Arg Asn Met Tyr Asp Ala Gly Ala Arg Val 850 855 860Gin Phe Glu Glu Gin lie Arg Asn Met Tyr Asp Ala Gly Ala Arg Val 850 855 860

Phe Leu Glu Phe Gly Pro Arg Gin Val Leu Ala Lys Leu lie Ala Glu 865 870 875 880Phe Leu Glu Phe Gly Pro Arg Gin Val Leu Ala Lys Leu lie Ala Glu 865 870 875 880

Met Phe Pro Ser Cys Thr Ala lie Ser Val Asn Pro Ala Ser Ser Gly 885 890 895Met Phe Pro Ser Cys Thr Ala lie Ser Val Asn Pro Ala Ser Ser Gly 885 890 895

Asp Ser Asp Val Gin Leu Arg Leu Ala Ala Val Lys Phe Ala Val Ser 900 905 910Asp Ser Asp Val Gin Leu Arg Leu Ala Ala Val Lys Phe Ala Val Ser 900 905 910

Gly Ala Ala Leu Ser Thr Phe Asp Pro Trp Glu Tyr Arg Lys Pro Gin 915 920 925Gly Ala Ala Leu Ser Thr Phe Asp Pro Trp Glu Tyr Arg Lys Pro Gin 915 920 925

Asp Leu Leu lie Arg Lys Pro Arg Lys Thr Ala Leu Val Leu Ser Ala 930 935 940Asp Leu Leu lie Arg Lys Pro Arg Lys Thr Ala Leu Val Leu Ser Ala 930 935 940

Ala Thr Tyr Val Ser Pro Lys Thr Leu Ala Glu Arg Lys Lys Ala Met 945 950 955 960Ala Thr Tyr Val Ser Pro Lys Thr Leu Ala Glu Arg Lys Lys Ala Met 945 950 955 960

Glu Asp lie Lys Leu Val Ser lie Thr Pro Arg Asp Ser Met Val Ser 965 970 975 lie Gly Lys lie Ala Gin Glu Val Arg Thr Ala Lys Gin Pro Leu Glu 980 985 990Glu Asp lie Lys Leu Val Ser lie Thr Pro Arg Asp Ser Met Val Ser 965 970 975 lie Gly Lys lie Ala Gin Glu Val Arg Thr Ala Lys Gin Pro Leu Glu 980 985 990

Thr Glu 工le Arg Arg Leu Asn Lys Glu Leu Glu His Leu Lys Arg Glu 995 1000 1005Thr Glu worker Arg Arg Leu Asn Lys Glu Leu Glu His Leu Lys Arg Glu 995 1000 1005

Leu Ala Ala Ala Lys Ala Ser Val Lys Ser Ala Ser Lys Ser Ser 1010 1015 1020Leu Ala Ala Ala Lys Ala Ser Val Lys Ser Ala Ser Lys Ser Ser 1010 1015 1020

Lys Glu Arg Ser Val Leu Ser Lys His Arg Ala Leu Leu Gin Asn 1025 1030 1035 lie Leu Gin Asp Tyr Asp Asp Leu Arg Val Val Pro Phe Ala Val 1040 1045 1050Lys Glu Arg Ser Val Leu Ser Lys His Arg Ala Leu Leu Gin Asn 1025 1030 1035 lie Leu Gin Asp Tyr Asp Asp Leu Arg Val Val Pro Phe Ala Val 1040 1045 1050

Arg Ser Val Ala Val Asp Asn Thr Ala Pro Tyr Ala Asp Gin Val 1055 1060 1065Arg Ser Val Ala Val Asp Asn Thr Ala Pro Tyr Ala Asp Gin Val 1055 1060 1065

Ser Thr Pro Ala Ser Glu Arg Ser Ala Ser Pro Leu Phe Glu Lys 1070 1075 1080Ser Thr Pro Ala Ser Glu Arg Ser Ala Ser Pro Leu Phe Glu Lys 1070 1075 1080

Arg Ser Ser Val Ser Ser Ala Arg Leu Ala Glu Ala Glu Ala Ala 1085 1090 1095 88 201038734Arg Ser Ser Val Ser Ser Ala Arg Leu Ala Glu Ala Glu Ala Ala 1085 1090 1095 88 201038734

Val Leu 1100 Ser Val Leu Ala Asp 1105 Lys Thr Gly Tyr Asp 1110 Ser Ser Met lie Glu 1115 Met Asp Met Asp Leu 1120 Glu Ser Glu Leu Gly 1125 Val Asp Ser lie Lys 1130 Arg Val Glu lie Met 1135 Ser Glu Val Gin Thr 1140 Leu Leu Ser Val Glu 1145 Val Ser Asp Val Asp 1150 Ala Leu Ser Arg Thr 1155 Lys Thr Val Gly Asp 1160 Val lie Glu Ala Met 1165 Lys Leu Glu Leu Gly 1170 Gly Pro Gin Ο Gly Gin 1175 Thr Leu Thr Ala Glu 1180 Ser lie Arg Gin Pro 1185 Pro Val Ser Glu Pro 1190 Ala Val Pro Thr Ser 1195 Ser Ser Ser Ser lie 1200 Ala Asn Val Ser Ser 1205 Ala Arg Leu Ala Glu 1210 Ala Glu Ala Ala Val 1215 Leu Ser Val - Leu Ala 1220 Asp Lys Thr Gly Tyr 1225 Asp Ser Ser Met lie 1230 Glu Met Asp - Met Asp 1235 Leu Glu Ser Glu Leu 1240 Gly Val Asp Ser lie 1245 Lys Arg Val Glu lie 1250 Met Ser Glu Val Gin 1255 Thr Leu Leu Ser Val 1260 Glu Val Ser Asp Val 1265 Asp Ala Leu Ser Arg 1270 Thr Lys Thr Val Gly 1275 Asp Val lie ο Glu Ala 1280 Met Lys Leu Glu Leu 1285 Gly Gly Pro Gin Gly 1290 Gin Thr Leu Thr Ala 1295 Glu Ser lie Arg Gin 1300 Pro Pro Val Ser Glu 1305 Pro Ala Val Pro Thr 1310 Ser Ser Ser Ser Ser 1315 lie Ala Asn Val Ser 1320 Ser Ala Arg Leu Ala 1325 Glu Ala Glu Ala Ala 1330 Val Leu Ser Val Leu 1335 Ala Asp Lys Thr Gly 1340 Tyr Asp Ser Ser Met 1345 lie Glu Met Asp Met 1350 Asp Leu Glu Ser Glu 1355 Leu Gly Val Asp Ser 1360 lie Lys Arg Val Glu 1365 lie Met Ser Glu Val Gin Thr Leu Leu Ser Val Glu Val Ser Asp Val Asp Ala 89 201038734 1370 1375 1380Val Leu 1100 Ser Val Leu Ala Asp 1105 Lys Thr Gly Tyr Asp 1110 Ser Ser Met lie Glu 1115 Met Asp Met Asp Leu 1120 Glu Ser Glu Leu Gly 1125 Val Asp Ser lie Lys 1130 Arg Val Glu lie Met 1135 Ser Glu Val Gin Thr 1140 Leu Leu Ser Val Glu 1145 Val Ser Asp Val Asp 1150 Ala Leu Ser Arg Thr 1155 Lys Thr Val Gly Asp 1160 Val lie Glu Ala Met 1165 Lys Leu Glu Leu Gly 1170 Gly Pro Gin Ο Gly Gin 1175 Thr Leu Thr Ala Glu 1180 Ser lie Arg Gin Pro 1185 Pro Val Ser Glu Pro 1190 Ala Val Pro Thr Ser 1195 Ser Ser Ser Ser lie 1200 Ala Asn Val Ser Ser 1205 Ala Arg Leu Ala Glu 1210 Ala Glu Ala Ala Val 1215 Leu Ser Val - Leu Ala 1220 Asp Lys Thr Gly Tyr 1225 Asp Ser Ser Met lie 1230 Glu Met Asp - Met Asp 1235 Leu Glu Ser Glu Leu 1240 Gly Val Asp Ser lie 1245 Lys Arg Val Glu lie 1250 Met Ser Glu Val Gin 1255 Thr Leu Leu Ser Val 1260 Glu Val Ser Asp Val 1265 Asp Ala Leu Ser Arg 1270 Thr Lys Thr Val Gly 1275 Asp Val lie ο Glu Ala 1280 Met Lys Leu Glu Leu 1285 Gly Gly Pro Gin Gly 1290 Gin Thr Leu Thr Ala 1295 Glu Ser lie Arg Gin 1300 Pro Pro Val Ser Glu 1305 Pro Ala Val Pro Thr 1310 Ser Ser Ser Ser Ser 1315 lie Ala Asn Val Ser 1320 Ser Ala Arg Leu Ala 1325 Glu Ala Glu Ala Ala 1330 Val Leu Ser Val Leu 1335 Ala Asp Lys Thr Gly 1340 Tyr Asp Ser Ser Met 1345 lie Glu Met Asp Met 1350 Asp Leu Glu Ser Glu 1355 Leu Gly Val Asp Ser 1360 lie Lys Arg Val Glu 1365 lie Met Ser Glu Val Gin Thr Leu Leu Ser Val Glu Val Ser Asp Val Asp Ala 89 201038734 1370 1375 1380

Leu Ser Arg Thr Lys Thr Val Gly Asp Val lie Glu Ala Met Lys 1385 1390 1395Leu Ser Arg Thr Lys Thr Val Gly Asp Val lie Glu Ala Met Lys 1385 1390 1395

Leu Glu Leu Gly Gly Pro Gin Gly Gin Thr Leu Thr Ala Glu Ser 1400 1405 1410 lie Arg Gin Pro Pro Val Ser Glu Pro Ala Val Pro Thr Ser Ser 1415 1420 1425Leu Glu Leu Gly Gly Gin Gin Gin Thr Leu Thr Ala Glu Ser 1400 1405 1410 lie Arg Gin Pro Pro Val Ser Glu Pro Ala Val Pro Thr Ser Ser 1415 1420 1425

Ser Ser Ser lie Ala Asn Val Leu Ser Ala Arg Leu Ala Glu Ala 1430 1435 1440Ser Ser Ser lie Ala Asn Val Leu Ser Ala Arg Leu Ala Glu Ala 1430 1435 1440

Glu Ala Ala Val Leu Ser Val Leu Ala Asp Lys Thr Gly Tyr Asp 1445 1450 1455Glu Ala Ala Val Leu Ser Val Leu Ala Asp Lys Thr Gly Tyr Asp 1445 1450 1455

Ser Ser Met lie Glu Met Asp Met Asp Leu Glu Ser Glu Leu GlySer Ser Met lie Glu Met Asp Met Asp Leu Glu Ser Glu Leu Gly

1460 1465 14701460 1465 1470

Val Asp Ser lie Lys Arg Val Glu lie Met Ser Glu Val Gin Thr 1475 1480 1485Val Asp Ser lie Lys Arg Val Glu lie Met Ser Glu Val Gin Thr 1475 1480 1485

Leu Leu Ser Val Glu Val Ser Asp Val Asp Ala Leu Ser Arg Thr 1490 1495 1500Leu Leu Ser Val Glu Val Ser Asp Val Asp Ala Leu Ser Arg Thr 1490 1495 1500

Lys Thr Val Gly Asp Val lie Glu Ala Met Lys Leu Glu Leu Gly 1505 1510 1515Lys Thr Val Gly Asp Val lie Glu Ala Met Lys Leu Glu Leu Gly 1505 1510 1515

Gly Pro Gin Gly Gin Thr Leu Thr Ala Glu Ser lie Arg Gin Pro 1520 1525 1530Gly Pro Gin Gly Gin Thr Leu Thr Ala Glu Ser lie Arg Gin Pro 1520 1525 1530

Pro Val Ser Glu Pro Ala Val Pro Thr Ser Ser Ser Ser Ser lie 1535 1540 1545Pro Val Ser Glu Pro Ala Val Pro Thr Ser Ser Ser Ser lie 1535 1540 1545

Ala Asn Val Ser Ser Ala Arg Leu Ala Glu Ala Glu Ala Ala Val 1550 1555 1560Ala Asn Val Ser Ser Ala Arg Leu Ala Glu Ala Glu Ala Ala Val 1550 1555 1560

Leu Ser Val Leu Ala Asp Lys Thr Gly Tyr Asp Ser Ser Met lie 1565 1570 1575Leu Ser Val Leu Ala Asp Lys Thr Gly Tyr Asp Ser Ser Met lie 1565 1570 1575

Glu Met Asp Met Asp Leu Glu Ser Glu Leu Gly Val Asp Ser lie 1580 1585 1590Glu Met Asp Met Asp Leu Glu Ser Glu Leu Gly Val Asp Ser lie 1580 1585 1590

Lys Arg Val Glu lie Met Ser Glu Val Gin Thr Leu Leu Ser Val 1595 1600 1605Lys Arg Val Glu lie Met Ser Glu Val Gin Thr Leu Leu Ser Val 1595 1600 1605

Glu Val Ser Asp Val Asp Ala Leu Ser Arg Thr Lys Thr Val Gly 1610 1615 1620Glu Val Ser Asp Val Asp Ala Leu Ser Arg Thr Lys Thr Val Gly 1610 1615 1620

Asp Val lie Glu Ala Met Lys Leu Glu Leu Gly Gly Pro Gin Gly 1625 1630 1635Asp Val lie Glu Ala Met Lys Leu Glu Leu Gly Gly Pro Gin Gly 1625 1630 1635

Gin Thr Leu Thr Ser Glu Pro lie His Gin Pro Pro Val Ser Glu 1640 1645 1650 90 201038734Gin Thr Leu Thr Ser Glu Pro lie His Gin Pro Pro Val Ser Glu 1640 1645 1650 90 201038734

Pro Ala 1655 Val Pro Thr Ser Ser 1660 Ser Ser Ser lie Ala 1665 Asn Val Ser Ser Ala 1670 Arg Leu Ala Glu Ala 1675 Glu Ala Ala Val Leu 1680 Ser Val Leu Ala Asp 1685 Lys Thr Gly Tyr Asp 1690 Ser Ser Met lie Glu 1695 Met Asp Met Asp Leu 17 00 Glu Ser Glu Leu Gly 1705 Val Asp Ser lie Lys 1710 Arg Val Glu lie Met 1715 Ser Glu Val Gin Thr 1720 Leu Leu Ser Val Glu 1725 Val Ser Asp Val Asp 1730 Ala Leu Ser Arg Thr 1735 Lys Thr Val Gly Asp 1740 Val lie Glu Ala Met 1745 Lys Met Glu Leu Gly 1750 Gly Pro Gin Gly Gin 1755 Thr Leu Thr Ala Glu 1760 Ser He Arg Gin Pro 1765 Pro Val Ser Glu Pro 1770 Ala Val Pro Thr Ser 1775 Ser Ser Ser Ser lie 1780 Ala Asn Val Ser Ser 1785 Ala Arg Leu Ala Glu 1790 Ala Glu Ala Ala Val 1795 Leu Ser Val Leu Ala 1800 Asp Lys Thr Gly Tyr 1805 Asp Ser Ser Met lie 1810 Glu Met Asp Met Asp 1815 Leu Glu Ser Glu Leu 1820 Gly Val Asp Ser lie 1825 Lys Arg Val Glu lie 1830 Met Ser Glu Val Gin 1835 Ala Leu Leu Ser Val 1840 Glu Val Ser Asp Val 1845 Asp Ala Leu Ser Arg 1850 Thr Lys Thr Val Gly 1855 Asp Val lie Glu Ala I860 Met Lys Met Glu Leu 1865 Gly Gly Pro Gin Gly 1870 Gin Thr Leu Thr Ala 1875 Glu Ser lie Arg Glu 1880 Pro Pro Val Ser Glu 1885 Pro Ala Val Pro Thr 1890 Ser Ser Ser Ser Ser 1895 lie Ala Asn Val Ser 1900 Ser Ala Arg Leu Ala 1905 Glu Ala Glu Ala Ala 1910 Val Leu Ser Val Leu 1915 Ala Asp Lys Thr Gly 1920 Tyr Asp Ser Ser Met 1925 lie Glu Met Asp Met 1930 Asp Leu Glu Ser Glu 1935 Leu Gly Val 91 201038734Pro Ala 1655 Val Pro Thr Ser Ser 1660 Ser Ser Ser lie Ala 1665 Asn Val Ser Ser Ala 1670 Arg Leu Ala Glu Ala 1675 Glu Ala Ala Val Leu 1680 Ser Val Leu Ala Asp 1685 Lys Thr Gly Tyr Asp 1690 Ser Ser Met lie Glu 1695 Met Asp Met Asp Leu 17 00 Glu Ser Glu Leu Gly 1705 Val Asp Ser lie Lys 1710 Arg Val Glu lie Met 1715 Ser Glu Val Gin Thr 1720 Leu Leu Ser Val Glu 1725 Val Ser Asp Val Asp 1730 Ala Leu Ser Arg Thr 1735 Lys Thr Val Gly Asp 1740 Val lie Glu Ala Met 1745 Lys Met Glu Leu Gly 1750 Gly Pro Gin Gly Gin 1755 Thr Leu Thr Ala Glu 1760 Ser He Arg Gin Pro 1765 Pro Val Ser Glu Pro 1770 Ala Val Pro Thr Ser 1775 Ser Ser Ser Ser lie 1780 Ala Asn Val Ser Ser 1785 Ala Arg Leu Ala Glu 1790 Ala Glu Ala Ala Val 1795 Leu Ser Val Leu Ala 1800 Asp Lys Thr Gly Tyr 1805 Asp Ser Ser Met lie 1810 Glu Met Asp Met Asp 1815 Leu Glu Ser Glu Leu 1820 Gly Val Asp Ser lie 1825 Lys Arg Val Glu lie 1830 Met Ser Glu Val Gin 1835 Ala Leu Leu Ser Val 1840 Glu Val Ser Asp Val 1845 Asp Ala Leu Ser Arg 1850 Thr Lys Thr Val Gly 1855 Asp Val lie Glu Ala I860 Met Lys Met Glu Leu 1865 Gly Gly Pro Gin Gly 1870 Gin Thr Leu Thr Ala 1875 Glu Ser lie Arg Glu 1880 Pro Pro Val Ser Glu 1885 Pro Ala Val Pro Thr 1890 Ser Ser Ser Ser Ser 1895 lie Ala Asn Val Ser 1900 Ser Ala Arg Leu Ala 1905 Glu Ala Glu Ala Ala 1910 Val Leu Ser Val Leu 1915 Ala Asp Lys Thr Gly 1920 Tyr Asp Ser Ser Met 1925 lie Glu Met Asp Met 1930 Asp Leu Glu Ser Glu 1935 Leu Gly Val 91 201038734

Asp Ser lie Lys Arg Val Glu lie Met Ser Glu Val Gin Thr Leu 1940 1945 1950Asp Ser lie Lys Arg Val Glu lie Met Ser Glu Val Gin Thr Leu 1940 1945 1950

Leu Ser Val Glu Val Ser Asp Val Asp Ala Leu Ser Arg Thr Lys 1955 1960 1965Leu Ser Val Glu Val Ser Asp Val Asp Ala Leu Ser Arg Thr Lys 1955 1960 1965

Thr Val Gly Asp Val lie Glu Ala Met Lys Leu Glu Leu Gly Glu 1970 1975 1980Thr Val Gly Asp Val lie Glu Ala Met Lys Leu Glu Leu Gly Glu 1970 1975 1980

Ser Ser Ser lie Glu Thr Leu Asn Cys Thr Glu Val Glu His Thr 1985 1990 1995Ser Ser Ser lie Glu Thr Leu Asn Cys Thr Glu Val Glu His Thr 1985 1990 1995

Ser Tyr Lys Ser Val Lys Ala Ser Gly Cys Glu Asn Val Asp Thr 2000 2005 2010Ser Tyr Lys Ser Val Lys Ala Ser Gly Cys Glu Asn Val Asp Thr 2000 2005 2010

Arg Phe Ala Lys Val Val Gin lie Ser Leu Pro Ser Lys Leu Lys 2015 2020 2025Arg Phe Ala Lys Val Val Gin lie Ser Leu Pro Ser Lys Leu Lys 2015 2020 2025

Ser Thr Val Ser His Asp Arg Pro Val lie Val Val Asp Asp Gly 2030 2035 2040Ser Thr Val Ser His Asp Arg Pro Val lie Val Val Asp Asp Gly 2030 2035 2040

Thr Pro Leu Thr Thr Glu Leu Cys Lys lie Leu Gly Gly Asn lie 2045 2050 2055Thr Pro Leu Thr Thr Glu Leu Cys Lys lie Leu Gly Gly Asn lie 2045 2050 2055

Val Val Leu Ser Tyr Gin Gly Lys Pro Ala Gly Pro Arg Gly Val 2060 2065 2070Val Val Leu Ser Tyr Gin Gly Lys Pro Ala Gly Pro Arg Gly Val 2060 2065 2070

Glu Val Pro Asp Leu Ser Glu Glu Ala Leu lie Gin Ala Leu Ala 2075 2080 2085Glu Val Pro Asp Leu Ser Glu Glu Ala Leu lie Gin Ala Leu Ala 2075 2080 2085

Leu lie Arg Ser Thr Tyr Gly Val Pro lie Gly Phe lie Cys Gin 2090 2095 2100Leu lie Arg Ser Thr Tyr Gly Val Pro lie Gly Phe lie Cys Gin 2090 2095 2100

Gin Val Ser Asn Val Ser Thr Lys Ala Gin Leu Cys Trp Ala Leu 2105 2110 2115Gin Val Ser Asn Val Ser Thr Lys Ala Gin Leu Cys Trp Ala Leu 2105 2110 2115

Leu Ala Ala Lys His Leu Lys Lys Asp Leu Asn Ala Val Leu Pro 2120 2125 2130Leu Ala Ala Lys His Leu Lys Lys Asp Leu Asn Ala Val Leu Pro 2120 2125 2130

Asp Ser Arg Ser Phe Phe Val Gly Val Val Arg Leu Asn Gly Lys 2135 2140 2145Asp Ser Arg Ser Phe Phe Val Gly Val Val Arg Leu Asn Gly Lys 2135 2140 2145

Leu Gly Thr Phe Glu Asn lie Ser Asp Phe Ser Lys Phe Asp Leu 2150 2155 2160Leu Gly Thr Phe Glu Asn lie Ser Asp Phe Ser Lys Phe Asp Leu 2150 2155 2160

Thr Lys Ala Leu Asp Tyr Gly Gin Arg Gly Ser Leu Leu Gly Leu 2165 2170 2175Thr Lys Ala Leu Asp Tyr Gly Gin Arg Gly Ser Leu Leu Gly Leu 2165 2170 2175

Cys Lys Ser Leu Asp Leu Glu Trp Glu Gin Val Phe Cys Arg Gly 2180 2185 2190 工le Asp Leu Ala Cys Asp Leu Met Pro Leu Gin Ala Ala Arg lie 2195 2200 2205Cys Lys Ser Leu Asp Leu Glu Trp Glu Gin Val Phe Cys Arg Gly 2180 2185 2190 work le Asp Leu Ala Cys Asp Leu Met Pro Leu Gin Ala Ala Arg lie 2195 2200 2205

Leu Arg Asn Glu Leu Gin Cys Pro Asn Met Arg Leu Arg Glu Val 2210 2215 2220 92 201038734Leu Arg Asn Glu Leu Gin Cys Pro Asn Met Arg Leu Arg Glu Val 2210 2215 2220 92 201038734

Gly Tyr 2225 Asp lie Ser Gly Ala Arg Tyr Thr lie Ser Thr Asp Asp 2230 2235 Leu Leu 2240 Cys Gly Pro Ser Lys Ala Lys Val Glu Ala Ala Asp Leu 2245 2250 Phe Leu 2255 Val Thr Gly Gly Ala Arg Gly lie Thr Pro His Cys Val 2260 2265 Arg Glu 2270 工le Ala Ser Arg Ser Pro Gly Thr Thr Phe Val Leu Val 2275 2280 Gly Arg 2285 Ser Glu Met Ser Asp Glu Pro Asp Trp Ala Val Gly His 2290 2295 Tyr Asn 赢 2300 Lys Asp Leu Asp Gin Ser Thr Met Lys His Leu Lys Ala 2305 2310 Thr His 2315 Ala Ala Gly Gly Val Lys Pro Thr Pro Lys Ala His Arg 2320 2325 Ala Leu 2330 Val Asn Arg Val Thr Gly Ser Arg Glu Val Arg Glu Ser 2335 2340 ' Leu Arg - 2345 Ala lie Gin Glu Ala Gly Ala Asn Val Glu Tyr lie Ala 2350 2355 Cys Asp * 2360 Val Ser Asp Glu Asn Lys Val Arg Gin Leu Val Gin Arg 2365 2370 Val Glu 2375 Gin Lys Tyr Gly Cys Glu lie Thr Gly lie Trp His Ala 2380 2385 Ser Gly 2390 Val Leu Arg Asp Lys Leu Val Glu Gin Lys Thr Thr Asp 2395 2400 Asp Phe 2405 Glu Ala Val Phe Gly Thr Lys Val Thr Gly Leu Val Asn 2410 2415 He Val 2420 Ser Gin Val Asn Met Ser Lys Leu Arg His Phe lie Leu 2425 2430 Phe Ser 2435 Ser Leu Ala Gly Phe His Gly Asn Lys Gly Gin Thr Asp 2440 2445 Tyr Ala 2450 lie Ala Asn Glu Ala Leu Asn Lys lie Ala His Thr Leu 2455 2460 Ser Ala 2465 Phe Leu Pro Lys Leu Asn Ala Lys Val Leu Asp Phe Gly 2470 2475 Pro Trp 2480 Val Gly Ser Gly Met Val Thr Glu Thr Leu Glu Lys His 2485 2490 Phe Lys Ala Met Gly Val Gin Thr lie Pro Leu Glu Pro Gly Ala 93 201038734 2495 2500 2505Gly Tyr 2225 Asp lie Ser Gly Ala Arg Tyr Thr lie Ser Thr Asp Asp 2230 2235 Leu Leu 2240 Cys Gly Pro Ser Lys Ala Lys Val Glu Ala Ala Asp Leu 2245 2250 Phe Leu 2255 Val Thr Gly Gly Ala Arg Gly lie Thr Pro His Cys Val 2260 2265 Arg Glu 2270 work le Ala Ser Arg Ser Pro Gly Thr Thr Phe Val Leu Val 2275 2280 Gly Arg 2285 Ser Glu Met Ser Asp Glu Pro Asp Trp Ala Val Gly His 2290 2295 Tyr Asn Win 2300 Lys Asp Leu Asp Gin Ser Thr Met Lys His Leu Lys Ala 2305 2310 Thr His 2315 Ala Ala Gly Gly Val Lys Pro Thr Pro Lys Ala His Arg 2320 2325 Ala Leu 2330 Val Asn Arg Val Thr Gly Ser Arg Glu Val Arg Glu Ser 2335 2340 ' Leu Arg - 2345 Ala lie Gin Glu Ala Gly Ala Asn Val Glu Tyr lie Ala 2350 2355 Cys Asp * 2360 Val Ser Asp Glu Asn Lys Val Arg Gin Leu Val Gin Arg 2365 2370 Val Glu 2375 Gin Lys Tyr Gly Cys Glu lie Thr Gly lie Trp His Ala 2380 2385 Ser Gly 2390 Val Leu Arg Asp Lys Leu Val Glu Gin Lys Thr Thr Asp 2395 2400 Asp Phe 2405 Glu Ala Val Phe Gly Thr Lys Val Thr Gly Leu Val Asn 2410 2415 He Val 2420 Ser Gin Val Asn Met Ser Lys Leu Arg His Phe lie Leu 2425 2430 Phe Ser 2435 Ser Leu Ala Gly Phe His Gly Asn Lys Gly Gin Thr Asp 2440 2445 Tyr Ala 2450 lie Ala Asn Glu Ala Leu Asn Lys lie Ala His Thr Leu 2455 2460 Ser Ala 2465 Phe Leu Pro Lys Leu Asn Ala Lys Val Leu Asp Phe Gly 2470 2475 Pro Trp 2480 Val Gly Ser Gly Met Val Thr Glu Thr Leu Glu Lys His 2485 2490 Phe Lys Ala Met Gly Val Gin Thr lie Pro Leu Glu Pro Gly Ala 93 201038734 2495 2500 2505

Arg Thr Val Ala Gin lie lie Leu Ala Ser Ser Pro Pro Gin Ser 2510 2515 2520Arg Thr Val Ala Gin lie lie Leu Ala Ser Ser Pro Pro Gin Ser 2510 2515 2520

Leu Leu Gly Asn Trp Gly Phe Pro Ala Thr Lys Pro Leu Gin Arg 2525 2530 2535Leu Leu Gly Asn Trp Gly Phe Pro Ala Thr Lys Pro Leu Gin Arg 2525 2530 2535

Ser Asn Val Val Thr Gly Thr Leu Ser Pro Glu Glu lie Glu Phe 2540 2545 2550 lie Ala Asp His Lys lie Gin Gly Arg Lys Val Leu Pro Met Met 2555 2560 2565Ser Asn Val Val Thr Gly Thr Leu Ser Pro Glu Glu lie Glu Phe 2540 2545 2550 lie Ala Asp His Lys lie Gin Gly Arg Lys Val Leu Pro Met Met 2555 2560 2565

Ala Ala lie Gly Phe Met Ala Ser lie Ala Glu Gly Leu Tyr Pro 2570 2575 2580Ala Ala lie Gly Phe Met Ala Ser lie Ala Glu Gly Leu Tyr Pro 2570 2575 2580

Gly Tyr Asn Leu Gin Gly Val Glu Asn Ala Gin Leu Phe Gin Gly 2585 2590 2595Gly Tyr Asn Leu Gin Gly Val Glu Asn Ala Gin Leu Phe Gin Gly 2585 2590 2595

Leu Thr lie Asn Gin Glu Thr Lys Phe Gin lie Thr Leu lie Glu 2600 2605 2610Leu Thr lie Asn Gin Glu Thr Lys Phe Gin lie Thr Leu lie Glu 2600 2605 2610

Glu His Asn Ser Glu Glu Asn Leu Asp Val Leu Thr Ser Leu Gly 2615 2620 2625Glu His Asn Ser Glu Glu Asn Leu Asp Val Leu Thr Ser Leu Gly 2615 2620 2625

Val Met Leu Glu Ser Gly Lys Val Leu Pro Ala Tyr Arg Cys Val 2630 2635 2640Val Met Leu Glu Ser Gly Lys Val Leu Pro Ala Tyr Arg Cys Val 2630 2635 2640

Val Cys Leu Asn Thr Thr Gin Gin Gin Pro Lys Leu Ser Pro Lys 2645 2650 2655 lie Leu Asn Leu Glu Val Asp Pro Ala Cys Glu Val Asn Pro Tyr 2660 2665 2670Val Cys Leu Asn Thr Thr Gin Gin Gin Pro Lys Leu Ser Pro Lys 2645 2650 2655 lie Leu Asn Leu Glu Val Asp Pro Ala Cys Glu Val Asn Pro Tyr 2660 2665 2670

Asp Gly Lys Ser Leu Phe His Gly Pro Leu Leu Gin Phe Val Gin 2675 2680 2685Asp Gly Lys Ser Leu Phe His Gly Pro Leu Leu Gin Phe Val Gin 2675 2680 2685

Gin Val Leu His Ser Ser Thr Lys Gly Leu Val Ala Lys Cys Arg 2690 2695 2700Gin Val Leu His Ser Ser Thr Lys Gly Leu Val Ala Lys Cys Arg 2690 2695 2700

Ala Leu Pro lie Lys Glu Ala 工le Arg Gly Pro Phe lie Lys Gin 2705 2710 2715Ala Leu Pro lie Lys Glu Ala lev Gly Pro Phe lie Lys Gin 2705 2710 2715

Thr Leu His Asp Pro lie Leu Asp Asp Val lie Phe Gin Leu Met 2720 2725 2730Thr Leu His Asp Pro lie Leu Asp Asp Val lie Phe Gin Leu Met 2720 2725 2730

Leu Val Trp Cys Arg Asn Ala Leu Gly Ser Ala Ser Leu Pro Asn 2735 2740 2745Leu Val Trp Cys Arg Asn Ala Leu Gly Ser Ala Ser Leu Pro Asn 2735 2740 2745

Arg lie Glu Lys Met Ser Tyr Phe Gly Asn Val Ser Glu Gly Ser 2750 2755 2760Arg lie Glu Lys Met Ser Tyr Phe Gly Asn Val Ser Glu Gly Ser 2750 2755 2760

Thr Phe Phe Ala Ser Val Thr Pro Val Gly Pro Arg Val Pro Lys 2765 2770 2775 94 201038734Thr Phe Phe Ala Ser Val Thr Pro Val Gly Pro Arg Val Pro Lys 2765 2770 2775 94 201038734

Asp Pro Val lie Lys Met Gin Phe Leu Leu Gin Asp Glu Ser Gly 2780 2785 2790Asp Pro Val lie Lys Met Gin Phe Leu Leu Gin Asp Glu Ser Gly 2780 2785 2790

Asn Thr Phe Ser Ser Gly Glu Gly Ser Val Val Leu Ser Asp Glu 2795 2800 2805Asn Thr Phe Ser Ser Gly Glu Gly Ser Val Val Leu Ser Asp Glu 2795 2800 2805

Leu Val Phe 2810 <210〉 57 <211> 2059 <212> PRT <213> 裂殖壺菌物種 <400> 57Leu Val Phe 2810 <210> 57 <211> 2059 <212> PRT <213> Schizochytrium species <400> 57

Met Ala Ala Arg Asn Val Ser Ala Ala His Glu Met His Asp Glu Lys 15 10 15Met Ala Ala Arg Asn Val Ser Ala Ala His Glu Met His Asp Glu Lys 15 10 15

Arg lie Ala Val Val Gly Met Ala Val Gin Tyr Ala Gly Cys Lys Thr 20 25 30Arg lie Ala Val Val Gly Met Ala Val Gin Tyr Ala Gly Cys Lys Thr 20 25 30

Lys Asp Glu Phe Trp Glu Val Leu Met Asn Gly Lys Val Glu Ser Lys 35 40 45Lys Asp Glu Phe Trp Glu Val Leu Met Asn Gly Lys Val Glu Ser Lys 35 40 45

Val lie Ser Asp Lys Arg Leu Gly Ser Asn Tyr Arg Ala Glu His Tyr 50 55 60Val lie Ser Asp Lys Arg Leu Gly Ser Asn Tyr Arg Ala Glu His Tyr 50 55 60

Lys Ala Glu Arg Ser Lys Tyr Ala Asp Thr Phe Cys Asn Glu Thr Tyr 65 70 75 80Lys Ala Glu Arg Ser Lys Tyr Ala Asp Thr Phe Cys Asn Glu Thr Tyr 65 70 75 80

Gly Thr Leu Asp Glu Asn Glu 工le Asp Asn Glu His Glu Leu Leu Leu 85 90 95Gly Thr Leu Asp Glu Asn Glu work le Asp Asn Glu His Glu Leu Leu Leu 85 90 95

Asn Leu Ala Lys Gin Ala Leu Ala Glu Thr Ser Val Lys Asp Ser Thr 100 105 110Asn Leu Ala Lys Gin Ala Leu Ala Glu Thr Ser Val Lys Asp Ser Thr 100 105 110

Arg Cys Gly lie Val Ser Gly Cys Leu Ser Phe Pro Met Asp Asn Leu 115 120 125Arg Cys Gly lie Val Ser Gly Cys Leu Ser Phe Pro Met Asp Asn Leu 115 120 125

Gin Gly Glu Leu Leu Asn Val Tyr Gin Asn His Val Glu Lys Lys Leu 130 135 140Gin Gly Glu Leu Leu Asn Val Tyr Gin Asn His Val Glu Lys Lys Leu 130 135 140

Gly Ala Arg Val Phe Lys Asp Ala Ser His Trp Ser Glu Arg Glu Gin 145 150 155 160Gly Ala Arg Val Phe Lys Asp Ala Ser His Trp Ser Glu Arg Glu Gin 145 150 155 160

Ser Asn Lys Pro Glu Ala Gly Asp Arg Arg lie Phe Met Asp Pro Ala 165 170 175Ser Asn Lys Pro Glu Ala Gly Asp Arg Arg lie Phe Met Asp Pro Ala 165 170 175

Ser Phe Val Ala Glu Glu Leu Asn Leu Gly Ala Leu His Tyr Ser Val 180 185 190Ser Phe Val Ala Glu Glu Leu Asn Leu Gly Ala Leu His Tyr Ser Val 180 185 190

Asp Ala Ala Cys Ala Thr Ala Leu Tyr Val Leu Arg Leu Ala Gin Asp 195 200 205Asp Ala Ala Cys Ala Thr Ala Leu Tyr Val Leu Arg Leu Ala Gin Asp 195 200 205

His Leu Val Ser Gly Ala Ala Asp Val Met Leu Cys Gly Ala Thr Cys 210 215 220 95 201038734His Leu Val Ser Gly Ala Ala Asp Val Met Leu Cys Gly Ala Thr Cys 210 215 220 95 201038734

Leu Pro Glu Pro Phe Phe lie Leu Ser Gly Phe Ser Thr Phe Gin Ala 225 230 235 240Leu Pro Glu Pro Phe Phe lie Leu Ser Gly Phe Ser Thr Phe Gin Ala 225 230 235 240

Met Pro Val Gly Thr Gly Gin Asn Val Ser Met Pro Leu His Lys Asp 245 250 255Met Pro Val Gly Thr Gly Gin Asn Val Ser Met Pro Leu His Lys Asp 245 250 255

Ser Gin Gly Leu Thr Pro Gly Glu Gly Gly Ser lie Met Val Leu Lys 260 265 270Ser Gin Gly Leu Thr Pro Gly Glu Gly Gly Ser lie Met Val Leu Lys 260 265 270

Arg Leu Asp Asp Ala lie Arg Asp Gly Asp His lie Tyr Gly Thr Leu 275 280 285Arg Leu Asp Asp Ala lie Arg Asp Gly Asp His lie Tyr Gly Thr Leu 275 280 285

Leu Gly Ala Asn Val Ser Asn Ser Gly Thr Gly Leu Pro Leu Lys Pro 290 295 300Leu Gly Ala Asn Val Ser Asn Ser Gly Thr Gly Leu Pro Leu Lys Pro 290 295 300

Leu Leu Pro Ser Glu Lys Lys Cys Leu Met Asp Thr Tyr Thr Arg lie 305 310 315 320Leu Leu Pro Ser Glu Lys Lys Cys Leu Met Asp Thr Tyr Thr Arg lie 305 310 315 320

Asn Val His Pro His Lys lie Gin Tyr Val Glu Cys His Ala Thr Gly 325 330 335Asn Val His Pro His Lys lie Gin Tyr Val Glu Cys His Ala Thr Gly 325 330 335

Thr Pro Gin Gly Asp Arg Val Glu lie Asp Ala Val Lys Ala Cys Phe 340 345 350Thr Pro Gin Gly Asp Arg Val Glu lie Asp Ala Val Lys Ala Cys Phe 340 345 350

Glu Gly Lys Val Pro Arg Phe Gly Thr Thr Lys Gly Asn Phe Gly His 355 360 365Glu Gly Lys Val Pro Arg Phe Gly Thr Thr Lys Gly Asn Phe Gly His 355 360 365

Thr Leu Val Ala Ala Gly Phe Ala Gly Met Cys Lys Val Leu Leu Ser 370 375 380Thr Leu Val Ala Ala Gly Phe Ala Gly Met Cys Lys Val Leu Leu Ser 370 375 380

Met Lys His Gly lie lie Pro Pro Thr Pro Gly lie Asp Asp Glu Thr 385 390 395 400Met Lys His Gly lie lie Pro Pro Thr Pro Gly lie Asp Asp Glu Thr 385 390 395 400

Lys Met Asp Pro Leu Val Val Ser Gly Glu Ala lie Pro Trp Pro Glu 405 410 415Lys Met Asp Pro Leu Val Val Ser Gly Glu Ala lie Pro Trp Pro Glu 405 410 415

Thr Asn Gly Glu Pro Lys Arg Ala Gly Leu Ser Ala Phe Gly Phe Gly 420 425 430Thr Asn Gly Glu Pro Lys Arg Ala Gly Leu Ser Ala Phe Gly Phe Gly 420 425 430

Gly Thr Asn Ala His Ala Val Phe Glu Glu His Asp Pro Ser Asn Ala 435 440 445Gly Thr Asn Ala His Ala Val Phe Glu Glu His Asp Pro Ser Asn Ala 435 440 445

Ala Cys Thr Gly His Asp Ser lie Ser Ala Leu Ser Ala Arg Cys Gly 450 455 460Ala Cys Thr Gly His Asp Ser lie Ser Ala Leu Ser Ala Arg Cys Gly 450 455 460

Gly Glu Ser Asn Met Arg lie Ala 工le Thr Gly Met Asp Ala Thr Phe 465 470 475 480Gly Glu Ser Asn Met Arg lie Ala gong le Thr Gly Met Asp Ala Thr Phe 465 470 475 480

Gly Ala Leu Lys Gly Leu Asp Ala Phe Glu Arg Ala lie Tyr Thr Gly 485 490 495Gly Ala Leu Lys Gly Leu Asp Ala Phe Glu Arg Ala lie Tyr Thr Gly 485 490 495

Ala His Gly Ala lie Pro Leu Pro Glu Lys Arq Trp Arq Phe Leu Gly 500 505 510Ala His Gly Ala lie Pro Leu Pro Glu Lys Arq Trp Arq Phe Leu Gly 500 505 510

Lys Asp Lys Asp Phe Leu Asp Leu Cys Gly Val Lys Ala Thr Pro His 515 520 525 96 201038734Lys Asp Lys Asp Phe Leu Asp Leu Cys Gly Val Lys Ala Thr Pro His 515 520 525 96 201038734

Gly Cys Tyr lie Glu Asp Val Glu Val Asp Phe Gin Arg Leu Arg Thr 530 535 540Gly Cys Tyr lie Glu Asp Val Glu Val Asp Phe Gin Arg Leu Arg Thr 530 535 540

Pro 545 Met Thr Pro Glu Asp Met Leu Leu Pro Gin Gin Leu Leu Ala Val 550 555 560 Thr Thr lie Asp Arg Ala lie Leu Asp Ser Gly Met Lys Lys Gly Gly 565 570 575 Asn Val Ala Val Phe Val Gly Leu Gly Thr Asp Leu Glu Leu Tyr Arg 580 585 590 His Arg Ala Arg Val Ala Leu Lys Glu Arg Val Arg Pro Glu Ala Ser 595 600 605 Lys Lys Leu Asn Asp Met Met Gin Tyr lie Asn Asp Cys Gly Thr Ser 610 615 620 o V Thr 625 Ser Tyr Thr Ser Tyr lie Gly Asn Leu Val Ala Thr Arg Val Ser 630 635 640 Ser Gin Trp Gly Phe Thr Gly Pro Ser Phe Thr lie Thr Glu Gly Asn 645 650 655 Asn Ser Val Tyr Arg Cys Ala Glu Leu Gly Lys Tyr Leu Leu Glu Thr 660 665 670 ， Gly Glu Val Asp Gly Val Val Val Ala Gly Val Asp Leu Cys Gly Ser 675 680 685 Ala Glu Asn Leu Tyr Val Lys Ser Arg Arg Phe Lys Val Ser Thr Ser 690 695 700 Asp 7〇5 Thr Pro Arg Ala Ser Phe Asp Ala Ala Ala Asp Gly Tyr Phe Val 710 715 720 Gly 〇 Glu Gly Cys Gly Ala Phe Val Leu Lys Arg Glu Thr Ser Cys Thr 725 730 735 Lys Asp Asp Arg lie Tyr Ala Cys Met Asp Ala lie Val Pro Gly Asn 740 745 750 Val Pro Ser Ala Cys Leu Arg Glu Ala Leu Asp Gin Ala Arg Val Lys 755 760 765 Pro Gly Asp lie Glu Met Leu Glu Leu Ser Ala Asp Ser Ala Arq His 770 775 780 Leu 785 Lys Asp Pro Ser Val Leu Pro Lys Glu Leu Thr Ala Glu Glu Glu 790 795 800 lie Gly Gly Leu Gin Thr lie Leu Arg Asp Asp Asp Lys Leu Pro Arq 805 810 815 Asn Val Ala Thr Gly Ser Val Lys Ala Thr Val Gly Asp Thr Glv Tvr 820 825 830 97 201038734Pro 545 Met Thr Pro Glu Asp Met Leu Leu Pro Gin Gin Leu Leu Ala Val 550 555 560 Thr Thr lie Asp Arg Ala lie Leu Asp Ser Gly Met Lys Lys Gly Gly 565 570 575 Asn Val Ala Val Phe Val Gly Leu Gly Thr Asp Leu Glu Leu Tyr Arg 580 585 590 His Arg Ala Arg Val Ala Leu Lys Glu Arg Val Arg Pro Glu Ala Ser 595 600 605 Lys Lys Leu Asn Asp Met Met Gin Tyr lie Asn Asp Cys Gly Thr Ser 610 615 620 o V Thr 625 Ser Tyr Thr Ser Tyr lie Gly Asn Leu Val Ala Thr Arg Val Ser 630 635 640 Ser Gin Trp Gly Phe Thr Gly Pro Ser Phe Thr lie Thr Glu Gly Asn 645 650 655 Asn Ser Val Tyr Arg Cys Ala Glu Leu Gly Lys Tyr Leu Leu Glu Thr 660 665 670 , Gly Glu Val Asp Gly Val Val Val Ala Gly Val Asp Leu Cys Gly Ser 675 680 685 Ala Glu Asn Leu Tyr Val Lys Ser Arg Arg Phe Lys Val Ser Thr Ser 690 695 700 Asp 7〇5 Thr Pro Arg Ala Ser Phe Asp Ala Ala Ala Asp Gly Tyr Phe Val 710 715 720 Gly Glu Gly Cys Gly Ala Phe Val Leu Lys Arg Glu Thr Ser Cys Thr 725 730 735 Lys Asp Asp Arg lie Tyr Ala Cys Met Asp Ala lie Va l Pro Gly Asn 740 745 750 Val Pro Ser Ala Cys Leu Arg Glu Ala Leu Asp Gin Ala Arg Val Lys 755 760 Pro Gly Asp lie Glu Met Leu Glu Leu Ser Ala Asp Ser Ala Arq His 770 775 780 Leu 785 Lys Asp Pro Ser Val Leu Pro Lys Glu Leu Thr Ala Glu Glu Glu 790 795 800 lie Gly Gly Leu Gin Thr lie Leu Arg Asp Asp Asp Lys Leu Pro Arq 805 810 815 Asn Val Ala Thr Gly Ser Val Lys Ala Thr Val Gly Asp Thr Glv Tvr 820 825 830 97 201038734

Ala Ser Gly Ala Ala Ser Leu lie Lys Ala Ala Leu Cys lie Tyr Asn 835 840 845Ala Ser Gly Ala Ala Ser Leu lie Lys Ala Ala Leu Cys lie Tyr Asn 835 840 845

Arg Tyr Leu Pro Ser Asn Gly Asp Asp Trp Asp Glu Pro Ala Pro Glu 850 855 860Arg Tyr Leu Pro Ser Asn Gly Asp Asp Trp Asp Glu Pro Ala Pro Glu 850 855 860

Ala Pro Trp Asp Ser Thr Leu Phe Ala Cys Gin Thr Ser Arg Ala Trp 865 870 875 880Ala Pro Trp Asp Ser Thr Leu Phe Ala Cys Gin Thr Ser Arg Ala Trp 865 870 875 880

Leu Lys Asn Pro Gly Glu Arg Arg Tyr Ala Ala Val Ser Gly Val Ser 885 890 895Leu Lys Asn Pro Gly Glu Arg Arg Tyr Ala Ala Val Ser Gly Val Ser 885 890 895

Glu Thr Arg Ser Cys Tyr Ser Val Leu Leu Ser Glu Ala Glu Gly His 900 905 910Glu Thr Arg Ser Cys Tyr Ser Val Leu Leu Ser Glu Ala Glu Gly His 900 905 910

Tyr Glu Arg Glu Asn Arg lie Ser Leu Asp Glu Glu Ala Pro Lys Leu 915 920 925Tyr Glu Arg Glu Asn Arg lie Ser Leu Asp Glu Glu Ala Pro Lys Leu 915 920 925

lie Val Leu Arg Ala Asp Ser His Glu Glu lie Leu Gly Arg Leu Asp 930 935 940Lie Val Leu Arg Ala Asp Ser His Glu Glu lie Leu Gly Arg Leu Asp 930 935 940

Lys lie Arg Glu Arg Phe Leu Gin Pro Thr Gly Ala Ala Pro Arg Glu 945 950 955 960Lys lie Arg Glu Arg Phe Leu Gin Pro Thr Gly Ala Ala Pro Arg Glu 945 950 955 960

Ser Glu Leu Lys Ala Gin Ala Arg Arg lie Phe Leu Glu Leu Leu Gly 965 970 975Ser Glu Leu Lys Ala Gin Ala Arg Arg lie Phe Leu Glu Leu Leu Gly 965 970 975

Glu Thr Leu Ala Gin Asp Ala Ala Ser Ser Gly Ser Gin Lys Pro Leu 980 985 990Glu Thr Leu Ala Gin Asp Ala Ala Ser Ser Gly Ser Gin Lys Pro Leu 980 985 990

Ala Leu Ser Leu Val Ser Thr Pro Ser Lys Leu Gin Arg Glu Val Glu 995 1000 1005Ala Leu Ser Leu Val Ser Thr Pro Ser Lys Leu Gin Arg Glu Val Glu 995 1000 1005

Leu Ala Ala Lys Gly lie Pro Arg Cys Leu Lys Met Arg Arg Asp 1010 1015 1020Leu Ala Ala Lys Gly lie Pro Arg Cys Leu Lys Met Arg Arg Asp 1010 1015 1020

Trp Ser Ser Pro Ala Gly Ser Arg Tyr Ala Pro Glu Pro Leu Ala 1025 1030 1035Trp Ser Ser Pro Ala Gly Ser Arg Tyr Ala Pro Glu Pro Leu Ala 1025 1030 1035

Ser Asp Arg Val Ala Phe Met Tyr Gly Glu Gly Arg Ser Pro Tyr 1040 1045 1050Ser Asp Arg Val Ala Phe Met Tyr Gly Glu Gly Arg Ser Pro Tyr 1040 1045 1050

Tyr Gly lie Thr Gin Asp lie His Arg lie Trp Pro Glu Leu His 1055 1060 1065Tyr Gly lie Thr Gin Asp lie His Arg lie Trp Pro Glu Leu His 1055 1060 1065

Glu Val lie Asn Glu Lys Thr Asn Arg Leu Trp Ala Glu Gly Asp 1070 1075 1080Glu Val lie Asn Glu Lys Thr Asn Arg Leu Trp Ala Glu Gly Asp 1070 1075 1080

Arg Trp Val Met Pro Arg Ala Ser Phe Lys Ser Glu Leu Glu Ser 1085 1090 1095Arg Trp Val Met Pro Arg Ala Ser Phe Lys Ser Glu Leu Glu Ser 1085 1090 1095

Gin Gin Gin Glu Phe Asp Arg Asn Met lie Glu Met Phe Arg Leu 1100 1105 1110Gin Gin Gin Glu Phe Asp Arg Asn Met lie Glu Met Phe Arg Leu 1100 1105 1110

Gly lie Leu Thr Ser lie Ala Phe Thr Asn Leu Ala Arg Asp Val 98 201038734 1115 1120 1125Gly lie Leu Thr Ser lie Ala Phe Thr Asn Leu Ala Arg Asp Val 98 201038734 1115 1120 1125

Leu Asn lie Thr Pro Lys Ala Ala Phe Gly Leu Ser Leu Gly Glu 1130 1135 1140 lie Ser Met lie Phe Ala Phe Ser Lys Lys Asn Gly Leu lie Ser 1145 1150 1155Leu Asn lie Thr Pro Lys Ala Ala Phe Gly Leu Ser Leu Gly Glu 1130 1135 1140 lie Ser Met lie Phe Ala Phe Ser Lys Lys Asn Gly Leu lie Ser 1145 1150 1155

Asp Gin Leu Thr Lys Asp Leu Arg Glu Ser Asp Val Trp Asn Lys 1160 1165 1170Asp Gin Leu Thr Lys Asp Leu Arg Glu Ser Asp Val Trp Asn Lys 1160 1165 1170

Ala Leu Ala Val Glu Phe Asn Ala Leu Arg Glu Ala Trp Gly lie 1175 1180 1185Ala Leu Ala Val Glu Phe Asn Ala Leu Arg Glu Ala Trp Gly lie 1175 1180 1185

Pro Gin Ser Val Pro Lys Asp Glu Phe Trp Gin Gly Tyr 工le Val 1190 1195 1200 ❹Pro Gin Ser Val Pro Lys Asp Glu Phe Trp Gin Gly Tyr work le Val 1190 1195 1200 ❹

Arg Gly Thr Lys Gin Asp lie Glu Ala Ala lie Ala Pro Asp Ser 1205 1210 1215Arg Gly Thr Lys Gin Asp lie Glu Ala Ala lie Ala Pro Asp Ser 1205 1210 1215

Lys Tyr Val Arg Leu Thr lie lie Asn Asp Ala Asn Thr Ala Leu 1220 1225 1230 lie Ser Gly Lys Pro Asp Ala Cys Lys Ala Ala lie Ala Arg Leu 1235 1240 1245Lys Tyr Val Arg Leu Thr lie lie Asn Asp Ala Asn Thr Ala Leu 1220 1225 1230 lie Ser Gly Lys Pro Asp Ala Cys Lys Ala Ala lie Ala Arg Leu 1235 1240 1245

Gly Gly Asn lie Pro Ala Leu Pro Val Thr Gin Gly Met Cys Gly 1250 1255 1260Gly Gly Asn lie Pro Ala Leu Pro Val Thr Gin Gly Met Cys Gly 1250 1255 1260

His Cys Pro Glu Val Gly Pro Tyr Thr Lys Asp lie Ala Lys lie 1265 1270 1275His Cys Pro Glu Val Gly Pro Tyr Thr Lys Asp lie Ala Lys lie 1265 1270 1275

His Ala Asn Leu Glu Phe Pro Val Val Asp Gly Leu Asp Leu Trp 1280 1285 1290His Ala Asn Leu Glu Phe Pro Val Val Asp Gly Leu Asp Leu Trp 1280 1285 1290

Thr Thr lie Asn Gin Lys Arg Leu Val Pro Arg Ala Thr Gly Ala 1295 1300 1305Thr Thr lie Asn Gin Lys Arg Leu Val Pro Arg Ala Thr Gly Ala 1295 1300 1305

Lys Asp Glu Trp Ala Pro Ser Ser Phe Gly Glu Tyr Ala Gly Gin 1310 1315 1320Lys Asp Glu Trp Ala Pro Ser Ser Phe Gly Glu Tyr Ala Gly Gin 1310 1315 1320

Leu Tyr Glu Lys Gin Ala Asn Phe Pro Gin lie Val Glu Thr lie 1325 1330 1335Leu Tyr Glu Lys Gin Ala Asn Phe Pro Gin lie Val Glu Thr lie 1325 1330 1335

Tyr Lys Gin Asn Tyr Asp Val Phe Val Glu Val Gly Pro Asn Asn 1340 1345 1350Tyr Lys Gin Asn Tyr Asp Val Phe Val Glu Val Gly Pro Asn Asn 1340 1345 1350

His Arg Ser Thr Ala Val Arg Thr Thr Leu Gly Pro Gin Arg Asn 1355 1360 1365His Arg Ser Thr Ala Val Arg Thr Thr Leu Gly Pro Gin Arg Asn 1355 1360 1365

His Leu Ala Gly Ala lie Asp Lys Gin Asn Glu Asp Ala Trp Thr 1370 1375 1380His Leu Ala Gly Ala lie Asp Lys Gin Asn Glu Asp Ala Trp Thr 1370 1375 1380

Thr lie Val Lys Leu Val Ala Ser Leu Lys Ala His Leu Val Pro 1385 1390 1395 99 201038734Thr lie Val Lys Leu Val Ala Ser Leu Lys Ala His Leu Val Pro 1385 1390 1395 99 201038734

Gly Val 1400Gly Val 1400

Thr lie Ser Pro Leu Tyr His Ser Lys Leu Val Ala Glu 1405 1410Thr lie Ser Pro Leu Tyr His Ser Lys Leu Val Ala Glu 1405 1410

Ala Gin 1415Ala Gin 1415

Ala Cys Tyr Ala Ala Leu Cys Lys Gly Glu Lys Pro Lys 1420 1425Ala Cys Tyr Ala Ala Leu Cys Lys Gly Glu Lys Pro Lys 1420 1425

Lys Asn 1430Lys Asn 1430

Lys Phe Val Arg Lys lie Gin Leu Asn Gly Arg Phe Asn 1435 1440Lys Phe Val Arg Lys lie Gin Leu Asn Gly Arg Phe Asn 1435 1440

Ser Lys 1445Ser Lys 1445

Ala Asp Pro lie Ser Ser Ala Asp Leu Ala Ser Phe Pro 1450 1455Ala Asp Pro lie Ser Ser Ala Asp Leu Ala Ser Phe Pro 1450 1455

Pro Ala 1460Pro Ala 1460

Asp Pro Ala lie Glu Ala Ala lie Ser Ser Arg lie Met 1465 1470Asp Pro Ala lie Glu Ala Ala lie Ser Ser Arg lie Met 1465 1470

Lys Pro 1475Lys Pro 1475

Val Ala Pro Lys Phe Tyr Ala Arg Leu Asn lie Asp Glu 1480 1485Val Ala Pro Lys Phe Tyr Ala Arg Leu Asn lie Asp Glu 1480 1485

Gin Asp 1490Gin Asp 1490

Glu Thr Arg Asp Pro lie Leu Asn Lys Asp Asn Ala Pro 1495 1500Glu Thr Arg Asp Pro lie Leu Asn Lys Asp Asn Ala Pro 1495 1500

Ser Ser 1505Ser Ser 1505

Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser 1510 1515Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser 1510 1515

Pro Ser 1520Pro Ser 1520

Pro Ala Pro Ser Ala Pro Val Gin Lys Lys Ala Ala Pro 1525 1530Pro Ala Pro Ser Ala Pro Val Gin Lys Lys Ala Ala Pro 1525 1530

Ala Ala 1535Ala Ala 1535

Glu Thr Lys Ala Val Ala Ser Ala Asp Ala Leu Arg Ser 1540 1545Glu Thr Lys Ala Val Ala Ser Ala Asp Ala Leu Arg Ser 1540 1545

Ala Leu 1550Ala Leu 1550

Leu Asp Leu Asp Ser Met Leu Ala Leu Ser Ser Ala Ser 1555 1560Leu Asp Leu Asp Ser Met Leu Ala Leu Ser Ser Ala Ser 1555 1560

Ala Ser 1565Ala Ser 1565

Gly Asn Leu Val Glu Thr Ala Pro Ser Asp Ala Ser Val 1570 1575Gly Asn Leu Val Glu Thr Ala Pro Ser Asp Ala Ser Val 1570 1575

He Val 1580He Val 1580

Pro Pro Cys Asn lie Ala Asp Leu Gly Ser Arg Ala Phe 1585 1590Pro Pro Cys Asn lie Ala Asp Leu Gly Ser Arg Ala Phe 1585 1590

Met Lys 1595Met Lys 1595

Thr Tyr Gly Val Ser Ala Pro Leu Tyr Thr Gly Ala Met 1600 1605Thr Tyr Gly Val Ser Ala Pro Leu Tyr Thr Gly Ala Met 1600 1605

Ala Lys 1610Ala Lys 1610

Gly lie Ala Ser Ala Asp Leu Val lie Ala Ala Gly Arg 1615 1620Gly lie Ala Ser Ala Asp Leu Val lie Ala Ala Gly Arg 1615 1620

Gin Gly 1625 lie Leu Ala Ser Phe Gly Ala Gly Gly Leu Pro Met Gin 1630 1635Gin Gly 1625 lie Leu Ala Ser Phe Gly Ala Gly Gly Leu Pro Met Gin 1630 1635

Val Val 1640Val Val 1640

Arg Glu Ser lie Glu Lys lie Gin Ala Ala Leu Pro Asn 1645 1650Arg Glu Ser lie Glu Lys lie Gin Ala Ala Leu Pro Asn 1645 1650

Gly Pro 1655Gly Pro 1655

Tyr Ala Val Asn Leu lie His Ser Pro Phe Asp Ser Asn 1660 1665Tyr Ala Val Asn Leu lie His Ser Pro Phe Asp Ser Asn 1660 1665

Leu Glu 1670Leu Glu 1670

Lys Gly Asn Val Asp Leu Phe Leu Glu Lys Gly Val Thr 1675 1680 100 201038734Lys Gly Asn Val Asp Leu Phe Leu Glu Lys Gly Val Thr 1675 1680 100 201038734

Phe Val Glu Ala Ser Ala Phe 1685 1690 Met Thr Leu Thr Pro Gin Val Val 1695 Arg Tyr Arg Ala Ala Gly Leu 1700 1705 Thr Arg Asn Ala Asp Gly Ser Val 1710 Asn lie Arg Asn Arg lie lie 1715 1720 Gly Lys Val Ser Arg Thr Glu Leu 1725 Ala Glu Met Phe Met Arq Pro 1730 1735 Ala Pro Glu His Leu Leu Gin Lys 1740 Leu lie Ala Ser Gly Glu lie 1745 1750 Asn Gin Glu Gin Ala Glu Leu Ala 1755 Arg Arg Val Pro Val Ala Asp 1760 1765 Asp lie Ala Val Glu Ala Asp Ser 1770 Gly Gly His Thr Asp Asn Arg 1775 1780 Pro lie His Val lie Leu Pro Leu 1785 He lie Asn Leu Arg Asp Arg 1790 1795 Leu His Arg Glu Cys Gly Tyr Pro 1800 Ala Asn Leu Arg Val Arg Val * 1805 1810 Gly Ala Gly Gly Gly He Gly Cys 1815 ， Pro Gin Ala Ala Leu Ala Thr 1820 1825 Phe Asn Met Gly Ala Ser Phe lie 1830 Val Thr Gly Thr Val Asn Gin 1835 1840 Val Ala Lys Gin Ser Gly Thr Cys 1845 Asp Asn Val Arg Lys Gin Leu 1850 1855 Ala Lys Ala Thr Tyr Ser Asp Val 1860 Cys Met Ala Pro Ala Ala Asp Q 1865 1870 Met Phe Glu Glu Gly Val Lys Leu 1875 Gin Val Leu Lys Lys Gly Thr 1880 1885 Met Phe Pro Ser Arg Ala Asn Lys 1890 Leu Tyr Glu Leu Phe Cys Lys 1895 1900 Tyr Asp Ser Phe Glu Ser Met Pro 1905 Pro Ala Glu Leu Ala Arg Val 1910 1915 Glu Lys Arg lie Phe Ser Arg Ala 1920 Leu Glu Glu Val Trp Asp Glu 1925 1930 Thr Lys Asn Phe Tyr lie Asn Arg 1935 Leu His Asn Pro Glu Lys lie 1940 1945 Gin Arg Ala Glu Arg Asp Pro Lys 1950 Leu Lys Met Ser Leu Cys Phe 1955 I960 Arg Trp Tyr Leu Ser Leu Ala Ser 1965 101 201038734Phe Val Glu Ala Ser Ala Phe 1685 1690 Met Thr Leu Thr Pro Gin Val Val 1695 Arg Tyr Arg Ala Ala Gly Leu 1700 1705 Thr Arg Asn Ala Asp Gly Ser Val 1710 Asn lie Arg Asn Arg lie lie 1715 1720 Gly Lys Val Ser Arg Thr Glu Leu 1725 Ala Glu Met Phe Met Arq Pro 1730 1735 Ala Pro Glu His Leu Leu Gin Lys 1740 Leu lie Ala Ser Gly Glu lie 1745 1750 Asn Gin Glu Gin Ala Glu Leu Ala 1755 Arg Arg Val Pro Val Ala Asp 1760 1765 Asp Lie Ala Val Glu Ala Asp Ser 1770 Gly Gly His Thr Asp Asn Arg 1775 1780 Pro lie His Val lie Leu Pro Leu 1785 He lie Asn Leu Arg Asp Arg 1790 1795 Leu His Arg Glu Cys Gly Tyr Pro 1800 Ala Asn Leu Arg Val Arg Val * 1805 1810 Gly Ala Gly Gly Gly He Gly Cys 1815 , Pro Gin Ala Ala Leu Ala Thr 1820 1825 Phe Asn Met Gly Ala Ser Phe lie 1830 Val Thr Gly Thr Val Asn Gin 1835 1840 Val Ala Lys Gin Ser Gly Thr Cys 1845 Asp Asn Val Arg Lys Gin Leu 1850 1855 Ala Lys Ala Thr Tyr Ser Asp Val 1860 Cys Met Ala Pro Ala Ala Asp Q 1865 1870 Met Phe Glu Glu Gly Val Lys Leu 1875 Gin Va l Leu Lys Lys Gly Thr 1880 1885 Met Phe Pro Ser Arg Ala Asn Lys 1890 Leu Tyr Glu Leu Phe Cys Lys 1895 1900 Tyr Asp Ser Phe Glu Ser Met Pro 1905 Pro Ala Glu Leu Ala Arg Val 1910 1915 Glu Lys Arg lie Phe Ser Arg Ala 1920 Leu Glu Glu Val Trp Asp Glu 1925 1930 Thr Lys Asn Phe Tyr lie Asn Arg 1935 Leu His Asn Pro Glu Lys lie 1940 1945 Gin Arg Ala Glu Arg Asp Pro Lys 1950 Leu Lys Met Ser Leu Cys Phe 1955 I960 Arg Trp Tyr Leu Ser Leu Ala Ser 1965 101 201038734

Arg Trp Ala Asn Thr Gly Ala Ser Asp Arg Val Met Asp Tyr Gin 1970 1975 1980Arg Trp Ala Asn Thr Gly Ala Ser Asp Arg Val Met Asp Tyr Gin 1970 1975 1980

Val Trp Cys Gly Pro Ala lie Gly Ser Phe Asn Asp Phe lie Lys 1985 1990 1995Val Trp Cys Gly Pro Ala lie Gly Ser Phe Asn Asp Phe lie Lys 1985 1990 1995

Gly Thr Tyr Leu Asp Pro Ala Val Ala Asn Glu Tyr Pro Cys Val 2000 2005 2010Gly Thr Tyr Leu Asp Pro Ala Val Ala Asn Glu Tyr Pro Cys Val 2000 2005 2010

Val Gin 工le Asn Lys Gin lie Leu Arg Gly Ala Cys Phe Leu Arg 2015 2020 2025Val Gin work Le Asn Lys Gin lie Leu Arg Gly Ala Cys Phe Leu Arg 2015 2020 2025

Arg Leu Glu lie Leu Arg Asn Ala Arg Leu Ser Asp Gly Ala Ala 2030 2035 2040Arg Leu Glu lie Leu Arg Asn Ala Arg Leu Ser Asp Gly Ala Ala 2030 2035 2040

Ala Leu Val Ala Ser lie Asp Asp Thr Tyr Val Pro Ala Glu Lys 2045 2050 2055Ala Leu Val Ala Ser lie Asp Asp Thr Tyr Val Pro Ala Glu Lys 2045 2050 2055

Leu <210> 58 <211> 1935 <212> PRT <213> 破橐壺菌物種 <400> 58Leu <210> 58 <211> 1935 <212> PRT <213> Thrombosporium species <400>

Met Gin Leu Pro Pro Ala His Ser Ala Asp Glu Asn Arg lie Ala Val 1 5 10 15Met Gin Leu Pro Pro Ala His Ser Ala Asp Glu Asn Arg lie Ala Val 1 5 10 15

Val Gly Met Ala Val Lys Tyr Ala Gly Cys Asp Asn Lys Glu Glu Phe 20 25 30Val Gly Met Ala Val Lys Tyr Ala Gly Cys Asp Asn Lys Glu Glu Phe 20 25 30

Trp Lys Thr Leu Met Asn Gly Ser 工le Asn Thr Lys Ser lie Ser Ala 35 40 45Trp Lys Thr Leu Met Asn Gly Ser Le Asn Thr Lys Ser lie Ser Ala 35 40 45

Ala Arg Leu Gly Ser Asn Lys Arg Asp Glu His Tyr Val Pro Glu Arg 50 55 60Ala Arg Leu Gly Ser Asn Lys Arg Asp Glu His Tyr Val Pro Glu Arg 50 55 60

Ser Lys Tyr Ala Asp Thr Phe Cys Asn Glu Arg Tyr Gly Cys lie Gin 65 70 75 80Ser Lys Tyr Ala Asp Thr Phe Cys Asn Glu Arg Tyr Gly Cys lie Gin 65 70 75 80

Gin Gly Thr Asp Asn Glu His Asp Leu Leu Leu Gly Leu Ala Gin Glu 85 90 95Gin Gly Thr Asp Asn Glu His Asp Leu Leu Leu Gly Leu Ala Gin Glu 85 90 95

Ala Leu Ala Asp Ala Ala Gly Arg Met Glu Lys Gin Pro Ser Glu Ala 100 105 110Ala Leu Ala Asp Ala Ala Gly Arg Met Glu Lys Gin Pro Ser Glu Ala 100 105 110

Phe Asp Leu Glu Asn Thr Gly lie Val Ser Gly Cys Leu Ser Phe Pro 115 120 125Phe Asp Leu Glu Asn Thr Gly lie Val Ser Gly Cys Leu Ser Phe Pro 115 120 125

Met Asp Asn Leu Gin Gly Glu Leu Leu Asn Leu Tyr Gin Ser His Val 130 135 140Met Asp Asn Leu Gin Gly Glu Leu Leu Asn Leu Tyr Gin Ser His Val 130 135 140

Glu Lys Gin Leu Pro Pro Ser Ala Leu Val Glu Ala Val Lys Leu Trp 145 150 155 160 102 201038734Glu Lys Gin Leu Pro Pro Ser Ala Leu Val Glu Ala Val Lys Leu Trp 145 150 155 160 102 201038734

Ser Glu Arg Gin Lys Ser Thr Lys Ala His Ala Gly Asp Lys Ara Ara 165 170 175Ser Glu Arg Gin Lys Ser Thr Lys Ala His Ala Gly Asp Lys Ara Ara 165 170 175

Phe 工le Asp Pro Ala Ser Phe Val Ala Asp Lys Leu Asn Leu Gly Pro 180 185 190Phe work le Asp Pro Ala Ser Phe Val Ala Asp Lys Leu Asn Leu Gly Pro 180 185 190

Leu His Tyr Ala He Asp Ala Ala Cys Ala Ser Ala Leu Tyr Val Leu 195 200 205Leu His Tyr Ala He Asp Ala Ala Cys Ala Ser Ala Leu Tyr Val Leu 195 200 205

Lys Leu Ala Gin Asp His Leu Val Ser Gly Ala Val Asp Met Met Leu 210 215 220Lys Leu Ala Gin Asp His Leu Val Ser Gly Ala Val Asp Met Met Leu 210 215 220

Cys Gly Ala Thr Cys Phe Pro Glu Pro Phe Phe lie Leu Ser Gly Phe 225 230 235 240Cys Gly Ala Thr Cys Phe Pro Glu Pro Phe Phe lie Leu Ser Gly Phe 225 230 235 240

Ser Thr Phe Gin Ala Met Pro Val Gly Ala Asp Gly Val Ser Leu Pro 245 250 255Ser Thr Phe Gin Ala Met Pro Val Gly Ala Asp Gly Val Ser Leu Pro 245 250 255

〇〇

Leu His Lys Thr Ser Ala Gly Leu Thr Pro Gly Glu Gly Gly Ser He 260 265 270Leu His Lys Thr Ser Ala Gly Leu Thr Pro Gly Glu Gly Gly Ser He 260 265 270

Met Val Leu Lys Arg Leu Lys Asp Ala lie Arg Asp Gly Asn His lie 275 280 285Met Val Leu Lys Arg Leu Lys Asp Ala lie Arg Asp Gly Asn His lie 275 280 285

Tyr Gly Val Leu Leu Glu Ala Asn Leu Ser Asn Ala Gly Cys Gly Leu 290 295 300Tyr Gly Val Leu Leu Glu Ala Asn Leu Ser Asn Ala Gly Cys Gly Leu 290 295 300

Pro Leu Ser Pro His Leu Pro Ser Glu Glu Ser Cvs lie Arg Asp Thr 305 310 315 320Pro Leu Ser Pro His Leu Pro Ser Glu Glu Ser Cvs lie Arg Asp Thr 305 310 315 320

Tyr Arg Arg Ala Gly Val Ala Ala Asp Gin Ser He Gin Tyr He Glu 325 330 335Tyr Arg Arg Ala Gly Val Ala Ala Asp Gin Ser He Gin Tyr He Glu 325 330 335

Cys His Ala Thr Gly Thr Pro Arg Gly Asp Val Val Glu He Glu Ala 340 345 350Cys His Ala Thr Gly Thr Pro Arg Gly Asp Val Val Glu He Glu Ala 340 345 350

Val Glu Arg Val Phe Lys Lys Asn Val Pro Arg Leu Gly Ser Thr Lys 355 360 365Val Glu Arg Val Phe Lys Lys Asn Val Pro Arg Leu Gly Ser Thr Lys 355 360 365

Gly Asn Phe Gly His Ser Leu Val Ala Ala Gly Phe Ala Gly Met Ala 370 375 380Gly Asn Phe Gly His Ser Leu Val Ala Ala Gly Phe Ala Gly Met Ala 370 375 380

Lys Leu Leu Leu Ala Met Glu His Gly Val lie Pro Pro Thr Pro Gly 385 390 395 400Lys Leu Leu Leu Ala Met Glu His Gly Val lie Pro Pro Thr Pro Gly 385 390 395 400

Leu Asp Ala Ser Asn Gin Ala Ser Glu His Val Val Thr Lys Ala lie 405 410 415Leu Asp Ala Ser Asn Gin Ala Ser Glu His Val Val Thr Lys Ala lie 405 410 415

Thr Trp Pro Glu Thr His Gly Ala Pro Lys Arg Ala Gly Leu Ser Ala 420 425 430Thr Trp Pro Glu Thr His Gly Ala Pro Lys Arg Ala Gly Leu Ser Ala 420 425 430

Phe Gly Phe Gly Gly Thr Asn Ala His Ala Leu Phe Glu Glu Phe Asn 435 440 445Phe Gly Phe Gly Gly Thr Asn Ala His Ala Leu Phe Glu Glu Phe Asn 435 440 445

Ala Glu Gly lie Ser Tyr Arg Pro Gly Lys Pro Pro Val Glu Ser Asn 450 455 460 103 201038734Ala Glu Gly lie Ser Tyr Arg Pro Gly Lys Pro Pro Val Glu Ser Asn 450 455 460 103 201038734

Thr Arg Pro Ser Val Val lie Thr Gly Met Asp Cys Thr Phe Gly Ser 465 470 475 480Thr Arg Pro Ser Val Val lie Thr Gly Met Asp Cys Thr Phe Gly Ser 465 470 475 480

Leu Glu Gly lie Asp Ala Phe Glu Thr Ala Leu Tyr Glu Gly Arg Asp 485 490 495Leu Glu Gly lie Asp Ala Phe Glu Thr Ala Leu Tyr Glu Gly Arg Asp 485 490 495

Ala Ala Arg Asp Leu Pro Ala Lys Arg Trp Arg Phe Leu Gly Glu Asp 500 505 510Ala Ala Arg Asp Leu Pro Ala Lys Arg Trp Arg Phe Leu Gly Glu Asp 500 505 510

Leu Glu Phe Leu Arg Ala lie Arg Leu Lys Glu Lys Pro Arg Gly Cys 515 520 525Leu Glu Phe Leu Arg Ala lie Arg Leu Lys Glu Lys Pro Arg Gly Cys 515 520 525

Phe Val Glu Ser Val Asp Val Asn Phe Arg Arg Leu Lys Thr Pro Leu 530 535 540Phe Val Glu Ser Val Asp Val Asn Phe Arg Arg Leu Lys Thr Pro Leu 530 535 540

Thr Pro Glu Asp Met Leu Arg Pro Gin Gin Leu Leu Ala Val Ser Thr 545 550 555 560Thr Pro Glu Asp Met Leu Arg Pro Gin Gin Leu Leu Ala Val Ser Thr 545 550 555 560

Met Asp Arg Ala lie 工le Asp Ala Gly Leu Lys Lys Gly Gin His Val 565 570 575Met Asp Arg Ala lie l Asp Ala Gly Leu Lys Lys Gly Gin His Val 565 570 575

Ala Val Leu Val Gly Leu Gly Thr Asp Leu Glu Leu Tyr Arg His Arg 580 585 590Ala Val Leu Val Gly Leu Gly Thr Asp Leu Glu Leu Tyr Arg His Arg 580 585 590

Ala Arg Val Ala Leu Lys Glu Val Leu His Pro Ser Leu Lys Ser Asp 595 600 605Ala Arg Val Ala Leu Lys Glu Val Leu His Pro Ser Leu Lys Ser Asp 595 600 605

Thr Ala 工le Leu Gin Lys 工le Met Gin Tyr Val Asn Asp Ala Gly Thr 610 615 620Thr Ala Le Le Gin Lys Le Met Gin Tyr Val Asn Asp Ala Gly Thr 610 615 620

Ser Thr Ser Tyr Thr Ser Tyr lie Gly Asn Leu Val Ala Thr Arg lie 625 630 635 640Ser Thr Ser Tyr Thr Ser Tyr lie Gly Asn Leu Val Ala Thr Arg lie 625 630 635 640

Ser Ser Gin Trp Gly Phe Thr Gly Pro Ser Phe Thr Val Thr Glu Gly 645 650 655Ser Ser Gin Trp Gly Phe Thr Gly Pro Ser Phe Thr Val Thr Glu Gly 645 650 655

Asn Asn Ser Val Tyr Arg Cys Ala Gin Leu Ala Lys Asp Met Leu Gin 660 665 670Asn Asn Ser Val Tyr Arg Cys Ala Gin Leu Ala Lys Asp Met Leu Gin 660 665 670

Val Asn Arg Val Asp Ala Val Val lie Ala Gly Val Asp Leu Asn Gly 675 680 685Val Asn Arg Val Asp Ala Val Val lie Ala Gly Val Asp Leu Asn Gly 675 680 685

Ser Ala Glu Ser Phe Phe Val Arg Ala Asn Arg Gin Lys lie Ser Lys 690 695 700Ser Ala Glu Ser Phe Phe Val Arg Ala Asn Arg Gin Lys lie Ser Lys 690 695 700

Leu Ser His Pro Cys Ala Ser Phe Asp Arg Asp Ala Asp Gly Phe Phe 705 710 715 720Leu Ser His Pro Cys Ala Ser Phe Asp Arg Asp Ala Asp Gly Phe Phe 705 710 715 720

Ala Gly Glu Gly Cys Gly Ala Leu Val Phe Lys Arg Leu Glu Asp Cys 725 730 735Ala Gly Glu Gly Cys Gly Ala Leu Val Phe Lys Arg Leu Glu Asp Cys 725 730 735

Ala Pro Gin Glu Lys lie Tyr Ala Ser lie Asp Ser lie Ala 工le Asp 740 745 750Ala Pro Gin Glu Lys lie Tyr Ala Ser lie Asp Ser lie Ala gong le Asp 740 745 750

Lys Glu Pro Thr Ser Ser Ala Val Lys Ala Val Tyr Gin Ser Asp Ser 104 201038734 755 760 765 Ser Leu Ser Asp lie 770 Glu Leu Leu Glu lie Ser Gly Asp Ser Lys Arg 775 780 Phe Ala Ala Phe Glu 785 Gly Ala Val Glu lie Gin Ser Ser Val Glu Ala 790 795 800 Gin Leu Lys Gly Leu 805 Ser Lys Val Leu Glu Pro Ala Lys Gly Gin Gly 810 815 Val Ala Val Gly Ser 820 Thr Arg Ala Thr Val Gly Asp lie Gly Tyr Ala 825 830 Thr Gly Ala Ala Ser 835 Leu lie Lys Thr Ala Leu Cys Leu Tyr Asn Arg 840 845 Tyr Leu Pro Ala Leu O 850 Ala Asn Trp Ser Gly Pro Cys Glu Gin Ser Ala 855 860 Trp Gly Ser Asn Met 865 Phe Val Cys His Glu Thr Arg Pro Trp Met Lys 870 875 880 Asn Gin Asn Glu Lys 885 Arg Cys Ala Leu lie Ser Gly Thr Asp Pro Ser 890 895 - His Thr Cys Phe Ser - 900 Leu Val Leu Ser Asp Thr Gly Cys Tyr Glu Glu 905 910 • His Asn Arg Thr Cys 915 Phe Asp Val Gin Ala Pro Gin Leu Val Leu lie 920 925 His Gly Phe Asp Gly 930 Lys Thr lie Val Arg Arg Leu Glu Gly Tyr Leu 935 940 Leu Glu Leu Val Glu 945 Gly His Ala Ser Pro Ser Glu Tyr Phe His Lys 950 955 960 Leu lie Gly Gin Ser 965 Leu Leu Glu Asn Ser Lys Glu Ser Lys Leu Thr 970 975 Leu Ser Leu Val Cys 980 Asn Pro Asn Gin Leu Gin Lys Glu Leu Met Leu 985 990 Ala lie Lys Gly Val 995 Gin Arg Ser Met Leu Thr Gly Lys Asp Trp Val 1000 1005Lys Glu Pro Thr Ser Ser Ala Val Lys Ala Val Tyr Gin Ser Asp Ser 104 201038734 755 760 765 Ser Leu Ser Asp lie 770 Glu Leu Leu Glu lie Ser Gly Asp Ser Lys Arg 775 780 Phe Ala Ala Phe Glu 785 Gly Ala Val Glu Lie Gin Ser Ser Val Glu Ala 790 795 800 Gin Leu Lys Gly Leu 805 Ser Lys Val Leu Glu Pro Ala Lys Gly Gin Gly 810 815 Val Ala Val Gly Ser 820 Thr Arg Ala Thr Val Gly Asp lie Gly Tyr Ala 825 830 Thr Gly Ala Ala Ser 835 Leu lie Lys Thr Ala Leu Cys Leu Tyr Asn Arg 840 845 Tyr Leu Pro Ala Leu O 850 Ala Asn Trp Ser Gly Pro Cys Glu Gin Ser Ala 855 860 Trp Gly Ser Asn Met 865 Phe Val Cys His Glu Thr Arg Pro Trp Met Lys 870 875 880 Asn Gin Asn Glu Lys 885 Arg Cys Ala Leu lie Ser Gly Thr Asp Pro Ser 890 895 - His Thr Cys Phe Ser - 900 Leu Val Leu Ser Asp Thr Gly Cys Tyr Glu Glu 905 910 • His Asn Arg Thr Cys 915 Phe Asp Val Gin Ala Pro Gin Leu Val Leu lie 920 925 His Gly Phe Asp Gly 930 Lys Thr lie Val Arg Arg Leu Glu Gly Tyr Leu 935 940 Leu Glu Leu Val Glu 945 Gly His Ala Ser Pro Ser Glu Tyr Phe His Lys 950 955 960 Leu lie Gly Gin Ser 965 Leu Leu Glu Asn Ser Lys Glu Ser Lys Leu Thr 970 975 Leu Ser Leu Val Cys 980 Asn Pro Asn Gin Leu Gin Lys Glu Leu Met Leu 985 990 Ala lie Lys Gly Val 995 Gin Arg Ser Met Leu Thr Gly Lys Asp Trp Val 1000 1005

Ser Pro Ser Gly Ser Cys Phe Ala Pro Asn Pro Leu Ser Ser Ala 1010 1015 1020Ser Pro Ser Gly Ser Cys Phe Ala Pro Asn Pro Leu Ser Ser Ala 1010 1015 1020

Lys Val Ala Phe Met Tyr Gly Glu Gly Arg Ser Pro Tyr Cys Gly 1025 1030 1035Lys Val Ala Phe Met Tyr Gly Glu Gly Arg Ser Pro Tyr Cys Gly 1025 1030 1035

Val Gly Leu Gly Leu His Arg Leu Trp Pro Gly Leu His Glu Asn 1040 1045 1050 105 201038734Val Gly Leu Gly Leu His Arg Leu Trp Pro Gly Leu His Glu Asn 1040 1045 1050 105 201038734

Val Asn Asn Lys Thr Val Asp Leu Trp Thr Glu Gly Asp Glv Trp 1055 1060 1065Val Asn Asn Lys Thr Val Asp Leu Trp Thr Glu Gly Asp Glv Trp 1055 1060 1065

Leu Tyr Pro Arg Thr Leu Thr Arg Glu Glu His Thr Lys Ala lie 1070 1075 1080Leu Tyr Pro Arg Thr Leu Thr Arg Glu Glu His Thr Lys Ala lie 1070 1075 1080

Glu Ser Phe Asn Ala Asn Gin lie Glu Met Phe Arg Ala Gly lie 1085 1090 1095Glu Ser Phe Asn Ala Asn Gin lie Glu Met Phe Arg Ala Gly lie 1085 1090 1095

Phe lie Ser Met Cys Gin Thr Asp Tyr Val Met Asn Val Leu Gly 1100 1105 1110Phe lie Ser Met Cys Gin Thr Asp Tyr Val Met Asn Val Leu Gly 1100 1105 1110

Val Gin Pro Lys Ala Gly Phe Gly Leu Ser Leu Gly Glu lie Ser 1115 1120 1125Val Gin Pro Lys Ala Gly Phe Gly Leu Ser Leu Gly Glu lie Ser 1115 1120 1125

Met Leu Phe Ala Met Ser Lys Glu Asn Cys Arg Gin Ser Gin Glu 1130 1135 1140Met Leu Phe Ala Met Ser Lys Glu Asn Cys Arg Gin Ser Gin Glu 1130 1135 1140

Met Thr Asn Arg Leu Arg Gly Ser Pro Val Trp Ser Asn Glu Leu 1145 1150 1155Met Thr Asn Arg Leu Arg Gly Ser Pro Val Trp Ser Asn Glu Leu 1145 1150 1155

Ala lie Asn Phe Asn Ala lie Arg Lys Leu Trp Lys lie Pro Arg 1160 1165 1170Ala lie Asn Phe Asn Ala lie Arg Lys Leu Trp Lys lie Pro Arg 1160 1165 1170

Gly Ala Pro Leu Glu Ser Phe Trp Gin Gly Tyr Leu Val His Gly 1175 1180 1185Gly Ala Pro Leu Glu Ser Phe Trp Gin Gly Tyr Leu Val His Gly 1175 1180 1185

Thr Arg Glu Glu Val Glu His Ala lie Gly Leu Ser Glu Pro Tyr 1190 1195 1200Thr Arg Glu Glu Val Glu His Ala lie Gly Leu Ser Glu Pro Tyr 1190 1195 1200

Val Arg Leu Leu lie Val Asn Asp Ser Arg Ser Ala Leu lie Ala 1205 1210 1215Val Arg Leu Leu lie Val Asn Asp Ser Arg Ser Ala Leu lie Ala 1205 1210 1215

Gly Lys Pro Asp Ala Cys Gin Ala Val lie Ser Arg Leu Asn Ser 1220 1225 1230Gly Lys Pro Asp Ala Cys Gin Ala Val lie Ser Arg Leu Asn Ser 1220 1225 1230

Lys Phe Pro Ser Leu Pro Val Lys Gin Gly Met lie Gly His Cys 1235 1240 1245Lys Phe Pro Ser Leu Pro Val Lys Gin Gly Met lie Gly His Cys 1235 1240 1245

Pro Glu Val Arg Ala Phe lie Lys Asp lie Gly Tyr lie His Glu 1250 1255 1260Pro Glu Val Arg Ala Phe lie Lys Asp lie Gly Tyr lie His Glu 1250 1255 1260

Thr Leu Arg lie Ser Asn Asp Tyr Ser Asp Cys Gin Leu Phe Ser 1265 1270 1275Thr Leu Arg lie Ser Asn Asp Tyr Ser Asp Cys Gin Leu Phe Ser 1265 1270 1275

Ala Val Thr Lys Gly Ala Leu Asp Ser Ser Thr Met Glu lie Lys 1280 1285 1290Ala Val Thr Lys Gly Ala Leu Asp Ser Ser Thr Met Glu lie Lys 1280 1285 1290

His Phe Val Gly Glu Val Tyr Ser Arg lie Ala Asp Phe Pro Gin 1295 1300 1305 lie Val Asn Thr Val His Ser Ala Gly Tyr Asp Val Phe Leu Glu 1310 1315 1320His Phe Val Gly Glu Val Tyr Ser Arg lie Ala Asp Phe Pro Gin 1295 1300 1305 lie Val Asn Thr Val His Ser Ala Gly Tyr Asp Val Phe Leu Glu 1310 1315 1320

Leu Gly Cys Asp Ala Ser Arg Ser Ala Ala Val Gin Asn lie Leu 1325 1330 1335 106 201038734Leu Gly Cys Asp Ala Ser Arg Ser Ala Ala Val Gin Asn lie Leu 1325 1330 1335 106 201038734

Gly Gly Gin Gly Lys Phe Leu Ser Thr Ala 工le Asp Lys Lys Glv 1340 1345 1350Gly Gly Gin Gly Lys Phe Leu Ser Thr Ala L. Asp Lys Lys Glv 1340 1345 1350

His Ser Ala Trp Ser Gin Val Leu Arg Ala Thr Ala Ser Leu Ala 1355 1360 1365His Ser Ala Trp Ser Gin Val Leu Arg Ala Thr Ala Ser Leu Ala 1355 1360 1365

Ala His Arg Val Pro Gly lie Ser lie Leu Asp Leu Phe His Pro 1370 1375 1380Ala His Arg Val Pro Gly lie Ser lie Leu Asp Leu Phe His Pro 1370 1375 1380

Asn Phe Arg Glu Met Cys Cys Thr Met Ala Thr Thr Pro Lys Val 1385 1390 1395Asn Phe Arg Glu Met Cys Cys Thr Met Ala Thr Thr Pro Lys Val 1385 1390 1395

Glu Asp Lys Phe Leu Arg Thr lie Gin lie Asn Gly Arg Phe Glu 1400 1405 1410Glu Asp Lys Phe Leu Arg Thr lie Gin lie Asn Gly Arg Phe Glu 1400 1405 1410

Lys Glu Met lie His Leu Glu Asp Thr Thr Leu Ser Cys Leu Pro 1415 1420 1425 ΟLys Glu Met lie His Leu Glu Asp Thr Thr Leu Ser Cys Leu Pro 1415 1420 1425 Ο

Ala Pro Ser Glu Ala Asn lie Ala Ala lie Gin Ser Arg Ser lie 1430 1435 1440Ala Pro Ser Glu Ala Asn lie Ala Ala lie Gin Ser Arg Ser lie 1430 1435 1440

Arg Ser Ala Ala Ala Arg Ser Gly Gin Ser His Asp Cys Ala Ser 1445 1450 1455Arg Ser Ala Ala Ala Arg Ser Gly Gin Ser His Asp Cys Ala Ser 1445 1450 1455

His Ser His Glu Glu Asn Lys Asp Ser Cys Pro Glu Lys Leu Lys 1460 1465 1470His Ser His Glu Glu Asn Lys Asp Ser Cys Pro Glu Lys Leu Lys 1460 1465 1470

Leu Asp Ser Val Ser Val Ala lie Asn Phe Asp Asn Asp Asp Arg 1475 1480 1485 工le Gin Leu Gly His Ala Gly Phe Arg Glu Met Tyr Asn Thr Arg 1490 1495 1500Leu Asp Ser Val Ser Val Ala lie Asn Phe Asp Asn Asp Asp Arg 1475 1480 1485 work le Gin Leu Gly His Ala Gly Phe Arg Glu Met Tyr Asn Thr Arg 1490 1495 1500

Tyr Ser Leu Tyr Thr Gly Ala Met Ala Lys Gly lie Ala Ser Ala 1505 1510 1515Tyr Ser Leu Tyr Thr Gly Ala Met Ala Lys Gly lie Ala Ser Ala 1505 1510 1515

Asp Leu Val lie Ala Ala Gly Lys Glu Gly lie Leu Ala Ser Tyr 1520 1525 1530Asp Leu Val lie Ala Ala Gly Lys Glu Gly lie Leu Ala Ser Tyr 1520 1525 1530

Gly Ala Gly Gly Leu Pro Leu Ala Thr Val Arg Lys Gly lie Asp 1535 1540 1545Gly Ala Gly Gly Leu Pro Leu Ala Thr Val Arg Lys Gly lie Asp 1535 1540 1545

Lys lie Gin Gin Ala Leu Pro Ser Gly Pro Tyr Ala Val Asn Leu 1550 1555 1560 lie His Ser Pro Phe Asp Gly Asn Leu Glu Gin Gly Asn Val Asp 1565 1570 1575Lys lie Gin Gin Ala Leu Pro Ser Gly Pro Tyr Ala Val Asn Leu 1550 1555 1560 lie His Ser Pro Phe Asp Gly Asn Leu Glu Gin Gly Asn Val Asp 1565 1570 1575

Leu Phe Leu Glu Lys Asn Val Arg Val Ala Glu Cys Ser Ala Phe 1580 1585 1590Leu Phe Leu Glu Lys Asn Val Arg Val Ala Glu Cys Ser Ala Phe 1580 1585 1590

Thr Thr Leu Thr Val Pro Val Val His Tyr Arg Ala Ala Gly Leu 1595 1600 1605Thr Thr Leu Thr Val Pro Val Val His Tyr Arg Ala Ala Gly Leu 1595 1600 1605

Val Arg Arg Gin Asp Gly Ser lie Leu lie Lys Asn Arg lie lie 1610 1615 1620 107 201038734Val Arg Arg Gin Asp Gly Ser lie Leu lie Lys Asn Arg lie lie 1610 1615 1620 107 201038734

Ala Lys Val Ser Arg Thr Glu Leu Ala Glu Met Phe Leu Arg Pro 1625 1630 1635Ala Lys Val Ser Arg Thr Glu Leu Ala Glu Met Phe Leu Arg Pro 1625 1630 1635

Ala Pro Gin lie lie Leu Glu Lys Leu Val Ala Ala Glu lie lie 1640 1645 1650Ala Pro Gin lie lie Leu Glu Lys Leu Val Ala Ala Glu lie lie 1640 1645 1650

Ser Ser Asp Gin Ala Arg Met Ala Ala Lys Val Pro Met Ala Asp 1655 1660 1665Ser Ser Asp Gin Ala Arg Met Ala Ala Lys Val Pro Met Ala Asp 1655 1660 1665

Asp lie Ala Val Glu Ala Asp Ser Gly Gly His Thr Asp Asn Arg 1670 1675 1680Asp lie Ala Val Glu Ala Asp Ser Gly Gly His Thr Asp Asn Arg 1670 1675 1680

Pro Met His Val 工le Leu Pro Leu lie lie Gin Leu Arg Asn Thr 1685 1690 1695 lie Leu Ala Glu Tyr Gly Cys Ala Thr Ala Phe Arg Thr Arg lie 1700 1705 1710Pro Met His Val Le Le Pro Leu lie lie Gin Leu Arg Asn Thr 1685 1690 1695 lie Leu Ala Glu Tyr Gly Cys Ala Thr Ala Phe Arg Thr Arg lie 1700 1705 1710

Gly Ala Gly Gly Gly lie Gly Cys Pro Ser Ala Ala Leu Ala Ala 1715 1720 1725Gly Ala Gly Gly Gly lie Gly Cys Pro Ser Ala Ala Lela Ala Ala 1715 1720 1725

Phe Asp Met Gly Ala Ser Phe Val Val Thr Gly Ser lie Asn Gin 1730 1735 1740 lie Cys Arg Glu Ala Gly Thr Cys Asp Thr Val Arg Glu Leu Leu 1745 1750 1755Phe Asp Met Gly Ala Ser Phe Val Val Thr Gly Ser lie Asn Gin 1730 1735 1740 lie Cys Arg Glu Ala Gly Thr Cys Asp Thr Val Arg Glu Leu Leu 1745 1750 1755

Ala Asn Ser Ser Tyr Ser Asp Val Thr Met Ala Pro Ala Ala Asp 1760 1765 1770Ala Asn Ser Ser Tyr Ser Asp Val Thr Met Ala Pro Ala Ala Asp 1760 1765 1770

Met Phe Asp Gin Gly Val Lys Leu Gin Val Leu Lys Arg Gly Thr 1775 1780 1785Met Phe Asp Gin Gly Val Lys Leu Gin Val Leu Lys Arg Gly Thr 1775 1780 1785

Met Phe Pro Ser Arg Ala Asn Lys Leu Arg Lys Leu Phe Val Asn 1790 1795 1800Met Phe Pro Ser Arg Ala Asn Lys Leu Arg Lys Leu Phe Val Asn 1790 1795 1800

Tyr Glu Ser Leu Glu Thr Leu Pro Ser Lys Glu Leu Lys Tyr Leu 1805 1810 1815Tyr Glu Ser Leu Glu Thr Leu Pro Ser Lys Glu Leu Lys Tyr Leu 1805 1810 1815

Glu Asn lie lie Phe Lys Gin Ala Val Asp Gin Val Trp Glu Glu 1820 1825 1830Glu Asn lie lie Phe Lys Gin Ala Val Asp Gin Val Trp Glu Glu 1820 1825 1830

Thr Lys Arg Phe Tyr Cys Glu Lys Leu Asn Asn Pro Asp Lys lie 1835 1840 1845Thr Lys Arg Phe Tyr Cys Glu Lys Leu Asn Asn Pro Asp Lys lie 1835 1840 1845

Ala Arg Ala Met Lys Asp Pro Lys Leu Lys Met Ser Leu Cys Phe 1850 1855 I860Ala Arg Ala Met Lys Asp Pro Lys Leu Lys Met Ser Leu Cys Phe 1850 1855 I860

Arg Trp Tyr Leu Ser Lys Ser Ser Gly Trp Ala Asn Ala Gly lie 1865 1870 1875Arg Trp Tyr Leu Ser Lys Ser Ser Gly Trp Ala Asn Ala Gly lie 1865 1870 1875

Lys Ser Arg Ala Leu Asp Tyr Gin lie Trp Cys Gly Pro Ala Met 1880 1885 1890Lys Ser Arg Ala Leu Asp Tyr Gin lie Trp Cys Gly Pro Ala Met 1880 1885 1890

Gly Ser Phe Asn Asn Phe Ala Ser Gly Thr Ser Leu Asp Trp Lys 108 201038734 1895 1900 1905Gly Ser Phe Asn Asn Phe Ala Ser Gly Thr Ser Leu Asp Trp Lys 108 201038734 1895 1900 1905

Val Thr Gly Val Phe Pro Gly Val Ala Glu Val Asn Met Ala lie 1910 1915 1920Val Thr Gly Val Phe Pro Gly Val Ala Glu Val Asn Met Ala lie 1910 1915 1920

Leu Asp Gly Ala Arg Glu Leu Ala Ala Lys Arg Asn 1925 1930 1935 <210> 59 <211> 2037 <212> PRT <213> 金黄色破囊壺菌 <400> 59Leu Asp Gly Ala Arg Glu Leu Ala Ala Lys Arg Asn 1925 1930 1935 <210> 59 <211> 2037 <212> PRT <213> Golden Thraustochyme <400> 59

Gin Ala lie Gly His Arg Ala Ala Arg Trp Ser Cys Arg Ser Lys Ser 15 10 15Gin Ala lie Gly His Arg Ala Ala Arg Trp Ser Cys Arg Ser Lys Ser 15 10 15

Lys Ala Arg Gly His Lys Ala Gin Lys Glu Met Asn Gin Gly Gly Arg 20 25 30Lys Ala Arg Gly His Lys Ala Gin Lys Glu Met Asn Gin Gly Gly Arg 20 25 30

Asn Asp Glu Gly Val Ser Val Ala Arg Ala Asp Pro Cys Pro Asp Thr 35 40 45Asn Asp Glu Gly Val Ser Val Ala Arg Ala Asp Pro Cys Pro Asp Thr 35 40 45

Arg lie Ala Val Val Gly Met Ala Val Glu Tyr Ala Gly Cys Arq Gly 50 55 60Arg lie Ala Val Val Gly Met Ala Val Glu Tyr Ala Gly Cys Arq Gly 50 55 60

Lys Glu Ala Phe Trp Asp Thr Leu Met Asn Gly Lys 工le Asn Ser Ala 65 70 75 80Lys Glu Ala Phe Trp Asp Thr Leu Met Asn Gly Lys work Le Asn Ser Ala 65 70 75 80

Cys lie Ser Asp Asp Arg Leu Gly Ser Ala Arg Arg Glu Glu His Tyr 85 90 95Cys lie Ser Asp Asp Arg Leu Gly Ser Ala Arg Arg Glu Glu His Tyr 85 90 95

Ala Pro Glu Arg Ser Lys Tyr Ala Asp Thr Phe Cys Asn Glu Arg Tyr 100 105 110Ala Pro Glu Arg Ser Lys Tyr Ala Asp Thr Phe Cys Asn Glu Arg Tyr 100 105 110

Gly Cys lie Asp Pro Lys Val Asp Asn Glu His Asp Leu Leu Leu Glv 115 120 125Gly Cys lie Asp Pro Lys Val Asp Asn Glu His Asp Leu Leu Leu Glv 115 120 125

Leu Ala Ala Ala Ala Leu Gin Asp Ala Gin Asp Arg Arg Ser Asp Gly 130 135 140Leu Ala Ala Ala Ala Leu Gin Asp Ala Gin Asp Arg Arg Ser Asp Gly 130 135 140

Gly Lys Phe Asp Pro Ala Gin Leu Lys Arq Cys Gly He Val Ser Glv 145 150 155 160Gly Lys Phe Asp Pro Ala Gin Leu Lys Arq Cys Gly He Val Ser Glv 145 150 155 160

Cys Leu Ser Phe Pro Met Asp Asn Leu Gin Gly Glu Leu Leu Asn Leu 165 170 175Cys Leu Ser Phe Pro Met Asp Asn Leu Gin Gly Glu Leu Leu Asn Leu 165 170 175

Tyr Gin Ala His Ala Glu Arg Arg He Gly Lys His Cys Phe Ala Asp 180 185 190Tyr Gin Ala His Ala Glu Arg Arg He Gly Lys His Cys Phe Ala Asp 180 185 190

Gin Thr Pro Trp Ser Thr Arg Thr Arg Ala Leu His Pro Leu Pro Gly 195 200 205Gin Thr Pro Trp Ser Thr Arg Thr Arg Ala Leu His Pro Leu Pro Gly 195 200 205

Asp Pro Arg Thr His Arg Asp Pro Ala Ser Phe Val Ala Gly Gin Leu 210 215 220 109 201038734Asp Pro Arg Thr His Arg Asp Pro Ala Ser Phe Val Ala Gly Gin Leu 210 215 220 109 201038734

Gly Leu Gly Pro Leu His Tyr Ser Leu Asp Ala Ala Cys Ala Ser Ala 225 230 235 240Gly Leu Gly Pro Leu His Tyr Ser Leu Asp Ala Ala Cys Ala Ser Ala 225 230 235 240

Leu Tyr Val Leu Arg Leu Ala Gin Asp His Leu Leu Ser Gly Glu Ala 245 250 255Leu Tyr Val Leu Arg Leu Ala Gin Asp His Leu Leu Ser Gly Glu Ala 245 250 255

Asp Leu Met Leu Cys Gly Ala Thr Cys Phe Pro Glu Pro Phe Phe lie 260 265 270Asp Leu Met Leu Cys Gly Ala Thr Cys Phe Pro Glu Pro Phe Phe lie 260 265 270

Leu Thr Gly Phe Ser Thr Phe His Ala Met Pro Val Gly Glu Asn Gly 275 280 285Leu Thr Gly Phe Ser Thr Phe His Ala Met Pro Val Gly Glu Asn Gly 275 280 285

a V r o e o S3 u e L u e L s • X H p s A g5 r 9 A 2 s 1 H e h p 〇 r p t e M r o e 9 s 2a V r o e o S3 u e L u e L s • X H p s A g5 r 9 A 2 s 1 H e h p 〇 r p t e M r o e 9 s 2

Gly Glu Ala AspGly Glu Ala Asp

Leu Met Leu Cys Gly Ala Thr Cys Phe Pro Glu Pro Phe Phe lie Leu 305 310 315 320Leu Met Leu Cys Gly Ala Thr Cys Phe Pro Glu Pro Phe Phe lie Leu 305 310 315 320

Thr Gly Phe Ser Thr Phe His Ala Met Pro Val Gly Glu Asn Gly Val 325 330 335Thr Gly Phe Ser Thr Phe His Ala Met Pro Val Gly Glu Asn Gly Val 325 330 335

ClCl

Ser Met Pro Phe His Arg Gin Pro Ser Glu Glu Ala Cys Leu Lys Ala 340 345 350 Thr Tyr Glu Leu Val Gly Val Pro Pro Arg Asp Val Gin Tyr Val Glu 355 360 365Ser Met Pro Phe His Arg Gin Pro Ser Glu Glu Ala Cys Leu Lys Ala 340 345 350 Thr Tyr Glu Leu Val Gly Val Pro Pro Arg Asp Val Gin Tyr Val Glu 355 360 365

Cys His Ala Thr Gly Thr Pro Gin Gly Asp Thr Val Glu Leu Gin Ala 370 375 380Cys His Ala Thr Gly Thr Pro Gin Gly Asp Thr Val Glu Leu Gin Ala 370 375 380

Val Lys Ala Cys Phe Glu Gly Ala Ser Pro Arg 385 390 395Val Lys Ala Cys Phe Glu Gly Ala Ser Pro Arg 385 390 395

s o yo L 4 r h T r e s y <—_ G e _—I Is o yo L 4 r h T r e s y <-_ G e _—I I

Gly Asn Phe Gly His Thr Leu Val Ala Ala Gly Phe Ala Gly Met Cys 405 410 415Gly Asn Phe Gly His Thr Leu Val Ala Ala Gly Phe Ala Gly Met Cys 405 410 415

Lys Val Leu Leu Ala Met Glu Arg Gly Val lie Pro Pro Thr Pro Gly 420 425 430 Val Asp Ser Gly Thr Gin lie Asp Pro Leu Val Val Thr Ala Ala Leu 435 440 445 〇Lys Val Leu Leu Ala Met Glu Arg Gly Val lie Pro Pro Pro Pro Gly 420 425 430 Val Asp Ser Gly Thr Gin lie Asp Pro Leu Val Val Thr Ala Ala Leu 435 440 445 〇

Pro Trp Pro Asp Thr Arg Gly Gly Pro Lys Arg Ala Gly Leu Ser Ala 450 455 460 Phe Gly Phe Gly Gly Thr Asn Ala His Ala Val Phe Glu Glu His He 465 470 475 480 Pro Ser Arg Ala Pro Pro Ala Val Leu Cys Gin Pro Arg Leu Gly Ser 485 490 495 Gly Pro Asn Arg Lys Leu Ala lie Val Gly Met Asp Ala Thr Phe Gly 500 505 510 Ser Leu Lys Gly Leu Ser Ala Leu Glu Ala Ala Leu Tyr Glu Ala Arg 515 520 525 110 201038734Pro Trp Pro Asp Thr Arg Gly Gly Pro Lys Arg Ala Gly Leu Ser Ala 450 455 460 Phe Gly Phe Gly Gly Thr Asn Ala His Ala Val Phe Glu Glu His He 465 470 475 480 Pro Ser Arg Ala Pro Pro Ala Val Leu Cys Gin Pro Arg Leu Gly Ser 485 490 495 Gly Pro Asn Arg Lys Leu Ala lie Val Gly Met Asp Ala Thr Phe Gly 500 505 510 Ser Leu Lys Gly Leu Ser Ala Leu Glu Ala Ala Leu Tyr Glu Ala Arg 515 520 525 110 201038734

His Ala Ala Arg Pro Leu Pro Ala Lys Arg Trp Arg Phe Leu Gly Glv 530 535 540His Ala Ala Arg Pro Leu Pro Ala Lys Arg Trp Arg Phe Leu Gly Glv 530 535 540

Asp Glu Ser Phe Leu His Glu lie Gly Leu Glu Cys Ser Pro His Gly 545 550 555 560Asp Glu Ser Phe Leu His Glu lie Gly Leu Glu Cys Ser Pro His Gly 545 550 555 560

Cys Tyr lie Glu Asp Val Asp Val Asp Phe Lys Arg Leu Arg Thr Pro 565 570 575Cys Tyr lie Glu Asp Val Asp Val Asp Phe Lys Arg Leu Arg Thr Pro 565 570 575

Met Val Pro Glu Asp Leu Leu Arg Pro Gin Gin Leu Leu Ala Val Ser 580 585 590Met Val Pro Glu Asp Leu Leu Arg Pro Gin Gin Leu Leu Ala Val Ser 580 585 590

Thr lie Asp Lys Ala lie Leu Asp Ser Gly Leu Ala Lys Gly Gly Asn 595 600 605Thr lie Asp Lys Ala lie Leu Asp Ser Gly Leu Ala Lys Gly Gly Asn 595 600 605

Val Ala Val Leu Val Gly Leu Gly Thr Asp Leu Glu Leu Tyr Arq His 610 615 620 ΟVal Ala Val Leu Val Gly Leu Gly Thr Asp Leu Glu Leu Tyr Arq His 610 615 620 Ο

Arg Ala Arg Val Ala Leu Lys Glu Arg Leu Gin Gly Leu Val Arq Ser 625 630 635 640Arg Ala Arg Val Ala Leu Lys Glu Arg Leu Gin Gly Leu Val Arq Ser 625 630 635 640

Ala Glu Gly Gly Ala Leu Thr Ser Arg Leu Met Asn Tyr lie Asn Asp 645 650 655Ala Glu Gly Gly Ala Leu Thr Ser Arg Leu Met Asn Tyr lie Asn Asp 645 650 655

Ser Gly Thr Ser Thr Ser Tyr Thr Ser Tyr lie Gly Asn Leu Val Ala 660 665 670Ser Gly Thr Ser Thr Ser Tyr Thr Ser Tyr lie Gly Asn Leu Val Ala 660 665 670

Thr Arg Val Ser Ser Gin Trp Gly Phe Thr Gly Pro Ser Phe Thr Val 675 680 685Thr Arg Val Ser Ser Gin Trp Gly Phe Thr Gly Pro Ser Phe Thr Val 675 680 685

Thr Glu Gly Ala Asn Ser Val His Arg Cys Ala Gin Leu Ala Lys Tyr 690 695 700Thr Glu Gly Ala Asn Ser Val His Arg Cys Ala Gin Leu Ala Lys Tyr 690 695 700

Met Leu Asp Arg Gly Glu Val Asp Ala Val Val Val Ala Gly Val Asp 705 710 715 720Met Leu Asp Arg Gly Glu Val Asp Ala Val Val Val Ala Gly Val Asp 705 710 715 720

Leu Cys Gly Ser Ala Glu Ala Phe Phe Val Arg Ser Arg Arg Met Gin 725 730 735 lie Ser Lys Ser Gin Arg Pro Ala Ala Pro Phe Asp Arg Ala Ala Asp 740 745 750Leu Cys Gly Ser Ala Glu Ala Phe Phe Val Arg Ser Arg Arg Met Gin 725 730 735 lie Ser Lys Ser Gin Arg Pro Ala Ala Pro Phe Asp Arg Ala Ala Asp 740 745 750

Gly Phe Phe Ala Gly Glu Gly Cys Gly Ala Leu Val Phe Lys Arg Leu 755 760 765Gly Phe Phe Ala Gly Glu Gly Cys Gly Ala Leu Val Phe Lys Arg Leu 755 760 765

Thr Asp Cys Val Ser Gly Glu Arg lie Tyr Ala Ser Leu Asp Ser Val 770 775 780Thr Asp Cys Val Ser Gly Glu Arg lie Tyr Ala Ser Leu Asp Ser Val 770 775 780

Val Val Ala Thr Thr Pro Arg Ala Ala Leu Arg Ala Ala Ala Gly Ser 785 790 795 800Val Val Ala Thr Thr Pro Arg Ala Ala Leu Arg Ala Ala Ala Gly Ser 785 790 795 800

Ala Arg Val Asp Pro Ala Ser 工le Asp Met Val Glu Leu Ser Ala Asp 805 810 815Ala Arg Val Asp Pro Ala Ser Le Asp Met Val Glu Leu Ser Ala Asp 805 810 815

Ser His Arg Phe Val Arg Ala Pro Gly Thr Val Ala Gin Pro Leu Thr 820 825 830 111 201038734Ser His Arg Phe Val Arg Ala Pro Gly Thr Val Ala Gin Pro Leu Thr 820 825 830 111 201038734

Ala Glu Val Glu Val Gly Ala Val Arg Glu Val lie Gly Thr Ala Gly 835 840 845Ala Glu Val Glu Val Gly Ala Val Arg Glu Val lie Gly Thr Ala Gly 835 840 845

Arg Gly Ser Arg Ser Val Ala Val Gly Ser Val Arg Ala Asn Val Gly 850 855 860Arg Gly Ser Arg Ser Val Ala Val Gly Ser Val Arg Ala Asn Val Gly 850 855 860

Asp Ala Gly Phe Ala Ser Gly Ala Ala Ala Leu Val Lys Thr Ala Leu 865 870 875 880Asp Ala Gly Phe Ala Ser Gly Ala Ala Ala Leu Val Lys Thr Ala Leu 865 870 875 880

Cys Leu His Asn Arg Tyr Leu Ala Ala Thr Pro Gly Trp Asp Ala Pro 885 890 895Cys Leu His Asn Arg Tyr Leu Ala Ala Thr Pro Gly Trp Asp Ala Pro 885 890 895

Ala Ala Gly Val Asp Phe Gly Ala Glu Leu Tyr Val Cys Arg Glu Ser 900 905 910Ala Ala Gly Val Asp Phe Gly Ala Glu Leu Tyr Val Cys Arg Glu Ser 900 905 910

Arg Ala Trp Val Lys Asn Ala Gly Val Ala Arg His Ala Ala lie Ser 915 920 925Arg Ala Trp Val Lys Asn Ala Gly Val Ala Arg His Ala Ala lie Ser 915 920 925

Gly Val Asp Glu Gly Gly Ser Cys Tyr Gly Leu Val Leu Ser Asp Val 930 935 940Gly Val Asp Glu Gly Gly Ser Cys Tyr Gly Leu Val Leu Ser Asp Val 930 935 940

Pro Gly Gin Tyr Glu Thr Gly Asn Arg lie Ser Leu Gin Ala Glu Ser 945 950 955 960Pro Gly Gin Tyr Glu Thr Gly Asn Arg lie Ser Leu Gin Ala Glu Ser 945 950 955 960

Pro Lys Leu Leu Leu Leu Ser Ala Pro Asp His Ala Ala Leu Leu Asp 965 970 975Pro Lys Leu Leu Leu Leu Ser Ala Pro Asp His Ala Ala Leu Leu Asp 965 970 975

Lys Val Ala Ala Glu Leu Ala Ala Leu Glu Gin Ala Asp Gly Leu Ser 980 985 990Lys Val Ala Ala Glu Leu Ala Ala Leu Glu Gin Ala Asp Gly Leu Ser 980 985 990

Ala Ala Ala Ala Ala Val Asp Arg Leu Leu Gly Glu Ser Leu Val Gly 995 1000 1005Ala Ala Ala Ala Ala Val Asp Arg Leu Leu Gly Glu Ser Leu Val Gly 995 1000 1005

Cys Ala Ala Gly Ser Gly Gly Leu Thr Leu Cys Leu Val Ala Ser 1010 1015 1020Cys Ala Ala Gly Ser Gly Gly Leu Thr Leu Cys Leu Val Ala Ser 1010 1015 1020

Pro Ala Ser Leu His Lys Glu Leu Ala Leu Ala His Arg Gly lie 1025 1030 1035Pro Ala Ser Leu His Lys Glu Leu Ala Leu Ala His Arg Gly lie 1025 1030 1035

Pro Arg Cys lie Lys Ala Arg Arg Asp Trp Ala Ser Pro Ala Gly 1040 1045 1050Pro Arg Cys lie Lys Ala Arg Arg Asp Trp Ala Ser Pro Ala Gly 1040 1045 1050

Ser Tyr Phe Ala Pro Glu Pro 工le Ala Ser Asp Arg Val Ala Phe 1055 1060 1065Ser Tyr Phe Ala Pro Glu Pro A Le Ser Asp Arg Val Ala Phe 1055 1060 1065

Met Tyr Gly Glu Gly Arg Ser Pro Tyr Cys Gly Val Gly Arg Asp 1070 1075 1080Met Tyr Gly Glu Gly Arg Ser Pro Tyr Cys Gly Val Gly Arg Asp 1070 1075 1080

Leu His Arg lie Trp Pro Ala Leu His Glu Arg Val Asn Ala Lys 1085 1090 1095Leu His Arg lie Trp Pro Ala Leu His Glu Arg Val Asn Ala Lys 1085 1090 1095

Thr Val Asn Leu Trp Gly Asp Gly Asp Ala Trp Leu Leu Pro Arq 1100 1105 1110Thr Val Asn Leu Trp Gly Asp Gly Asp Ala Trp Leu Leu Pro Arq 1100 1105 1110

Ala Thr Ser Ala Glu Glu Glu Glu Gin Leu Cys Arg Asn Phe Asp 112 201038734 1115 1120 1125Ala Thr Ser Ala Glu Glu Glu Glu Gin Leu Cys Arg Asn Phe Asp 112 201038734 1115 1120 1125

Ser Asn Gin Val Glu Met Phe 1130 1135 Arg Thr Gly Val Tyr lie Ser Met 1140 Cys Leu Thr Asp Leu Ala Arg 1145 1150 Ser Leu lie Gly Leu Gly Pro Lys 1155 Ala Ser Phe Gly Leu Ser Leu 1160 1165 Gly Glu Val Ser Met Leu Phe Ala 1170 Leu Ser Glu Ser Asn Cys Arg 1175 1180 Leu Ser Glu Glu Met Thr Arg Arg 1185 Leu Arg Ala Ser Pro Val Trp 1190 1195 Asn Ser Glu Leu Ala Val Glu Phe 1200 的 Asn Ala Leu Arg Lys Leu Trp 1205 1210 Gly Val Ala Pro Gly Ala Pro Val 1215 Asp Ser Phe Trp Gin Gly Tyr 1220 1225 Val Val Arg Ala Thr Arg Ala Gin 1230 Val Glu Gin Ala lie Gly Glu 1235 1240 Asp Asn Gin Phe Val Arg Leu Leu 1245 • lie Val Asn Asp Ser Gin Ser - 1250 1255 Val Leu lie Ala Gly Lys Pro Ala 1260 、 Ala Cys Glu Ala Val lie Ala 1265 1270 Arg lie Gly Ser lie Leu Pro Pro 1275 Leu Gin Val Ser Gin Gly Met 1280 1285 Val Gly His Cys Ala Glu Val Leu 1290 Pro Tyr Thr Ser Glu lie Gly 1295 1300 Arg lie His Asn Met Leu Arg Phe 1305 Pro Ser Gin Asp Glu Thr Gly 1310 1315 Gly Cys Lys Met Tyr Ser Ser Val 1320 Ser Asn Ser Arg lie Gly Pro 1325 1330 Val Glu Glu Ser Gin Met Gly Pro 1335 Gly Thr Glu Leu Val Phe Ser 1340 1345 Pro Ser Met Glu Asp Phe Val Ala 1350 Gin Leu Tyr Ser Arg Val Ala 1355 1360 Asp Phe Pro Ala lie Thr Glu Ala 1365 Val Tyr Gin Gin Gly His Asp 1370 1375 Val Phe Val Glu Val Gly Pro Asp 1380 His Ser Arg Ser Ala Ala Val 1385 1390 Arg Ser Thr Leu Gly Pro Thr Arg 1395 113 201038734Ser Asn Gin Val Glu Met Phe 1130 1135 Arg Thr Gly Val Tyr lie Ser Met 1140 Cys Leu Thr Asp Leu Ala Arg 1145 1150 Ser Leu lie Gly Leu Gly Pro Lys 1155 Ala Ser Phe Gly Leu Ser Leu 1160 1165 Gly Glu Val Ser Met Leu Phe Ala 1170 Leu Ser Glu Ser Asn Cys Arg 1175 1180 Leu Ser Glu Glu Met Thr Arg Arg 1185 Leu Arg Ala Ser Pro Val Trp 1190 1195 Asn Ser Glu Leu Ala Val Glu Phe 1200 Asn Ala Leu Arg Lys Leu Trp 1205 1210 Gly Val Ala Pro Gly Ala Pro Val 1215 Asp Ser Phe Trp Gin Gly Tyr 1220 1225 Val Val Arg Ala Thr Arg Ala Gin 1230 Val Glu Gin Ala lie Gly Glu 1235 1240 Asp Asn Gin Phe Val Arg Leu Leu 1245 • lie Val Asn Asp Ser Gin Ser - 1250 1255 Val Leu lie Ala Gly Lys Pro Ala 1260, Ala Cys Glu Ala Val lie Ala 1265 1270 Arg lie Gly Ser lie Leu Pro Pro 1275 Leu Gin Val Ser Gin Gly Met 1280 1285 Val Gly His Cys Ala Glu Val Leu 1290 Pro Tyr Thr Ser Glu lie Gly 1295 1300 Arg lie His Asm Met Leu Arg Phe 1305 Pro Ser Gin Asp Glu Thr Gly 1310 1315 Gly Cys Lys Met Tyr Ser Ser Val 1320 Ser Asn Ser Arg lie Gly Pro 1325 1330 Val Glu Glu Ser Gin Met Gly Pro 1335 Gly Thr Glu Leu Val Phe Ser 1340 1345 Pro Ser Met Glu Asp Phe Val Ala 1350 Gin Leu Tyr Ser Arg Val Ala 1355 1360 Asp Phe Pro Ala lie Thr Glu Ala 1365 Val Tyr Gin Gin Gly His Asp 1370 1375 Val Phe Val Glu Val Gly Pro Asp 1380 His Ser Arg Ser Ala Ala Val 1385 1390 Arg Ser Thr Leu Gly Pro Thr Arg 1395 113 201038734

Arg His lie Ala Val Ala Met Asp Arg Lys Gly Glu Ser Ala Trp 1400 1405 1410Arg His lie Ala Val Ala Met Asp Arg Lys Gly Glu Ser Ala Trp 1400 1405 1410

Ser Gin Leu Leu Lys Met Leu Ala Thr Leu Ala Ser His Arg Val 1415 1420 1425Ser Gin Leu Leu Lys Met Leu Ala Thr Leu Ala Ser His Arg Val 1415 1420 1425

Pro Gly Leu Asp Leu Ser Ser Met Tyr His Pro Ala Val Val Glu 1430 1435 1440Pro Gly Leu Asp Leu Ser Ser Met Tyr His Pro Ala Val Val Glu 1430 1435 1440

Arg Cys Arg Leu Ala Leu Ala Ala Gin Arg Ser Gly Gin Pro Glu 1445 1450 1455Arg Cys Arg Leu Ala Leu Ala Ala Gin Arg Ser Gly Gin Pro Glu 1445 1450 1455

Gin Arg Asn Lys Phe Leu Arg Thr lie Glu Val Asn Gly Phe Tyr 1460 1465 1470Gin Arg Asn Lys Phe Leu Arg Thr lie Glu Val Asn Gly Phe Tyr 1460 1465 1470

Asp Pro Ala Asp Ala Thr lie Pro Glu Ala Val Ala Thr lie Leu 1475 1480 1485Asp Pro Ala Asp Ala Thr lie Pro Glu Ala Val Ala Thr lie Leu 1475 1480 1485

Pro Ala Thr Ala Ala lie Ser Pro Pro Lys Leu Gly Ala Pro His 1490 1495 1500Pro Ala Thr Ala Ala lie Ser Pro Pro Lys Leu Gly Ala Pro His 1490 1495 1500

Asp Ser Gin Pro Glu Ala Glu Ala Arg Pro Val Gly Glu Ala Ser 1505 1510 1515Asp Ser Gin Pro Glu Ala Glu Ala Arg Pro Val Gly Glu Ala Ser 1505 1510 1515

Val Pro Arg Arg Ala Thr Ser Ser Ser Lys Leu Ala Arg Thr Leu 1520 1525 1530Val Pro Arg Arg Ala Thr Ser Ser Ser Lys Leu Ala Arg Thr Leu 1520 1525 1530

Ala lie Asp Ala Cys Asp Ser Asp Val Arg Ala Ala Leu Leu Asp 1535 1540 1545Ala lie Asp Ala Cys Asp Ser Asp Val Arg Ala Ala Leu Leu Asp 1535 1540 1545

Leu Asp Ala Pro He Ala Val Gly Gly Ser Ser Arg Ala Gin Val 1550 1555 1560Leu Asp Ala Pro He Ala Val Gly Gly Ser Ser Arg Ala Gin Val 1550 1555 1560

Pro Pro Cys Pro Val Ser Ala Leu Gly Ser Ala Ala Phe Arq Ala 1565 1570 1575Pro Pro Cys Pro Val Ser Ala Leu Gly Ser Ala Ala Phe Arq Ala 1565 1570 1575

Ala His Gly Val Asp Tyr Ala Leu Tyr Met Gly Ala Met Ala Lys 1580 1585 1590Ala His Gly Val Asp Tyr Ala Leu Tyr Met Gly Ala Met Ala Lys 1580 1585 1590

Gly Val Ala Ser Ala Glu Met Val lie Ala Ala Gly Lys Ala Arq 1595 1600 1605Gly Val Ala Ser Ala Glu Met Val lie Ala Ala Gly Lys Ala Arq 1595 1600 1605

Met Leu Ala Ser Phe Gly Ala Gly Gly Leu Pro Leu Gly Glu Val 1610 1615 1620Met Leu Ala Ser Phe Gly Ala Gly Gly Leu Pro Leu Gly Glu Val 1610 1615 1620

Glu Glu Ala Leu Asp Lys lie Gin Ala Ala Leu Pro Glu Glv Pro 1625 1630 1635Glu Glu Ala Leu Asp Lys lie Gin Ala Ala Leu Pro Glu Glv Pro 1625 1630 1635

Phe Ala Val Asn Leu lie His Ser Pro Phe Asp Pro Asn Leu Glu 1640 1645 1650Phe Ala Val Asn Leu lie His Ser Pro Phe Asp Pro Asn Leu Glu 1640 1645 1650

Glu Gly Asn Val Glu Leu Phe Leu Arg Arg Gly lie Arq Leu Val 1655 1660 1665Glu Gly Asn Val Glu Leu Phe Leu Arg Arg Gly lie Arq Leu Val 1655 1660 1665

Glu Ala Ser Ala Phe Met Ser Val Thr Pro Ser Leu Val Ara Tvr 1670 1675 1680 114Glu Ala Ser Ala Phe Met Ser Val Thr Pro Ser Leu Val Ara Tvr 1670 1675 1680 114

Arg Val Ala Gly Leu Glu 1685 Leu Asn Arg Val lie Gly 17 00 Met Phe Met Arg Pro Pro 1715 Ala Gin Gly Leu Val Thr 1730 Val Pro Leu Val Asp Asp 1745 His Thr Asp Asn Arg Pro 17 60 Asn Arg Val Arg Val Gly 1790 Ala Ala Arg Ala Ala Phe 1805 Gly Ser lie Asn Gin Leu 1820 Val Arg Ala Ala Leu Ala 1835 Ala Pro Ala Ala Asp Met 1850 Glu Leu Ala Arg Leu Glu 1895 Glu Val Trp Asn Glu Thr 1910 Asn Pro Ala Lys Val Ala 1925 Met Ser Leu Cys Phe Arg 1940 Ala Ser Thr Gly Gin Val 1955 Arg 1690 Gly Pro Gly Gly Thr 1695 Ala Arg Val Lys 1705 Val Ser Arg Ala Glu 1710 Leu Ala Glu Pro 1720 Ala Ala lie Val Ser 1725 Lys Leu Leu Glu 1735 Glu Gin Ala Ser Leu 1740 Ala Glu lie Val 1750 Ala lie Glu Ala Asp 1755 Ser Gly Gly lie 1765 His Val Val Leu Pro 1770 Val Val Leu Met 1780 Arg Glu Cys Lys Tyr 1785 Pro Ala Ala Ala 1795 Gly Gly Gly He Gly 1800 Cys Pro Ala Asp 1810 Met Gly Ala Ala Phe 1815 Val Leu Thr Thr 1825 Arg Gin Ala Gly Thr 1830 Ser Asp Ser Arg 1840 Ala Thr Tyr Ser Asp 1845 Val Thr Met Phe 1855 Asp Gin Gly Val Lys 1860 Leu Gin Val Phe 1870 Pro Ala Arg Ala Asn 1875 Lys Leu Tyr Gin 1885 Ser Leu Asp Ala lie 1890 Pro Arg Ala Lys 1900 Arg Val Phe Arg Met 1905 Ser lie Asp Lys 1915 Gin Phe Tyr Glu Thr 1920 Arg Leu Asn Arg 1930 Ala Glu Arg Asp Pro 1935 Lys Leu Lys Trp 1945 Tyr Leu Ser Lys Ser 1950 Ser Lys Trp Gly 1960 Arg Glu Leu Asp Tyr 1965 Gin Val Trp 201038734Arg Val Ala Gly Leu Glu 1685 Leu Asn Arg Val lie Gly 17 00 Met Phe Met Arg Pro Pro 1715 Ala Gin Gly Leu Val Thr 1730 Val Pro Leu Val Asp Asp 1745 His Thr Asp Asn Arg Pro 17 60 Asn Arg Val Arg Val Gly 1790 Ala Ala Arg Ala Ala Phe 1805 Gly Ser lie Asn Gin Leu 1820 Val Arg Ala Ala Leu Ala 1835 Ala Pro Ala Ala Asp Met 1850 Glu Leu Ala Arg Leu Glu 1895 Glu Val Trp Asn Glu Thr 1910 Asn Pro Ala Lys Val Ala 1925 Met Ser Leu Cys Phe Arg 1940 Ala Ser Thr Gly Gin Val 1955 Arg 1690 Gly Pro Gly Gly Thr 1695 Ala Arg Val Lys 1705 Val Ser Arg Ala Glu 1710 Leu Ala Glu Pro 1720 Ala Ala lie Val Ser 1725 Lys Leu Leu Glu 1735 Glu Gin Ala Ser Leu 1740 Ala Glu lie Val 1750 Ala lie Glu Ala Asp 1755 Ser Gly Gly lie 1765 His Val Val Leu Pro 1770 Val Val Leu Met 1780 Arg Glu Cys Lys Tyr 1785 Pro Ala Ala Ala 1795 Gly Gly Gly He Gly 1800 Cys Pro Ala Asp 1810 Met Gly Ala Ala Phe 1815 Val Leu Thr Thr 1825 Arg Gin Ala Gly Thr 1830 Ser Asp Ser Arg 1840 Ala Thr Tyr Ser Asp 1845 Val Thr Met Phe 1855 Asp Gin Gly Val Lys 1860 Leu Gin Val Phe 1870 Pro Ala Arg Ala Asn 1875 Lys Leu Tyr Gin 1885 Ser Leu Asp Ala lie 1890 Pro Arg Ala Lys 1900 Arg Val Phe Arg Met 1905 Ser lie Asp Lys 1915 Gin Phe Tyr Glu Thr 1920 Arg Leu Asn Arg 1930 Ala Glu Arg Asp Pro 1935 Lys Leu Lys Trp 1945 Tyr Leu Ser Lys Ser 1950 Ser Lys Trp Gly 1960 Arg Glu Leu Asp Tyr 1965 Gin Val Trp 201038734

177 5177 5

Leu Lys Arg Gly Thr Met 〇 1865Leu Lys Arg Gly Thr Met 〇 1865

Glu Leu Phe Thr Thr Tyr 1880 115 201038734Glu Leu Phe Thr Thr Tyr 1880 115 201038734

Cys Gly Pro Thr lie Gly Ala Phe Asn Glu Phe Val Lys Gly Ser 1970 1975 1980Cys Gly Pro Thr lie Gly Ala Phe Asn Glu Phe Val Lys Gly Ser 1970 1975 1980

Ser Leu Asp Ala Glu Ala Cys Gly Gly Arg Phe Pro Cys Val Val 1985 1990 1995Ser Leu Asp Ala Glu Ala Cys Gly Gly Arg Phe Pro Cys Val Val 1985 1990 1995

Arg Val Asn Gin Glu lie Leu Cys Gly Ala Ala Tyr Glu Gin Arg 2000 2005 2010Arg Val Asn Gin Glu lie Leu Cys Gly Ala Ala Tyr Glu Gin Arg 2000 2005 2010

Leu Ala Arg Phe Met Leu Leu Ala Gly Arg Glu Ser Ala Asp Ala 2015 2020 2025Leu Ala Arg Phe Met Leu Leu Ala Gly Arg Glu Ser Ala Asp Ala 2015 2020 2025

Leu Ala Tyr Thr Val Ala Glu Ala Arg 2030 2035 <210〉 60 <211> 1470 <212> PRT <213> 破囊壺菌物種 <400> 60Leu Ala Tyr Thr Val Ala Glu Ala Arg 2030 2035 <210> 60 <211> 1470 <212> PRT <213> Thraustochytrium Species <400> 60

Met Gly Pro Arg Val Ala Ser Gly Lys Val Pro Ala Trp Glu Met Ser 1 5 10 15Met Gly Pro Arg Val Ala Ser Gly Lys Val Pro Ala Trp Glu Met Ser 1 5 10 15

Lys Ser Glu Leu Cys Asp Asp Arg Thr Val Val Phe Asp Tyr Glu Glu 20 25 30Lys Ser Glu Leu Cys Asp Asp Arg Thr Val Val Phe Asp Tyr Glu Glu 20 25 30

Leu Leu Glu Phe Ala Glu Gly Asp He Ser Lys Val Phe Gly Pro Glu 35 40 45Leu Leu Glu Phe Ala Glu Gly Asp He Ser Lys Val Phe Gly Pro Glu 35 40 45

Phe Lys Val Val Asp Gly Phe Arg Arg Arg Val Arg Leu Pro Ala Arg 50 55 60Phe Lys Val Val Asp Gly Phe Arg Arg Arg Val Arg Leu Pro Ala Arg 50 55 60

Glu Tyr Leu Leu Val Thr Arg Val Thr Leu Met Asp Ala Glu Val Gly 65 70 75 80Glu Tyr Leu Leu Val Thr Arg Val Thr Leu Met Asp Ala Glu Val Gly 65 70 75 80

Asn Phe Arg Val Gly Ala Arg Met Val Thr Glu Tyr Asp Val Pro Val 85 90 95Asn Phe Arg Val Gly Ala Arg Met Val Thr Glu Tyr Asp Val Pro Val 85 90 95

Asn Gly Glu Leu Ser Glu Gly Gly Asp Val Pro Trp Ala Val Leu Val 100 105 110Asn Gly Glu Leu Ser Glu Gly Gly Asp Val Pro Trp Ala Val Leu Val 100 105 110

Glu Ala Gly Gin Cys Asp Leu Leu Leu lie Ser Tyr Met Gly lie Asp 115 120 125Glu Ala Gly Gin Cys Asp Leu Leu Leu lie Ser Tyr Met Gly lie Asp 115 120 125

Phe Gin Cys Lys Gly Glu Arg Val Tyr Arg Leu Leu Asn Thr Thr Leu 130 135 140Phe Gin Cys Lys Gly Glu Arg Val Tyr Arg Leu Leu Asn Thr Thr Leu 130 135 140

Thr Phe Phe Gly Val Ala Lys Glu Gly Glu Thr Leu Val Tyr Asp lie 145 150 155 160Thr Phe Phe Gly Val Ala Lys Glu Gly Glu Thr Leu Val Tyr Asp lie 145 150 155 160

Arg Val Thr Gly Phe Ala Lys Arg Pro Asp Gly Asp 165 170 t 5 e 7 Ml r e s e h pArg Val Thr Gly Phe Ala Lys Arg Pro Asp Gly Asp 165 170 t 5 e 7 Ml r e s e h p

Phe Phe Glu Tyr Asp Cys Tyr Cys Asn Gly Lys Leu Leu lie Glu Met 180 185 190 116 201038734Phe Phe Glu Tyr Asp Cys Tyr Cys Asn Gly Lys Leu Leu lie Glu Met 180 185 190 116 201038734

Arg Asp Gly Ser Ala Gly Phe Phe Thr Asp Glu Glu Leu Ala Ala Gly 195 200 205Arg Asp Gly Ser Ala Gly Phe Phe Thr Asp Glu Glu Leu Ala Ala Gly 195 200 205

Lys Gly Val Val Val Thr Arg Ala Gin Gin Asn Met Arg Asp Lys lie 210 215 220Lys Gly Val Val Val Thr Arg Ala Gin Gin Asn Met Arg Asp Lys lie 210 215 220

Val Arg Gin Ser lie Glu Pro Phe Ala Leu Ala Ala Cys Thr His Lys 225 230 235 240Val Arg Gin Ser lie Glu Pro Phe Ala Leu Ala Ala Cys Thr His Lys 225 230 235 240

Thr Thr Leu Asn Glu Ser Asp Met Gin Ser Leu Val Glu Arg Asn Trp 245 250 255Thr Thr Leu Asn Glu Ser Asp Met Gin Ser Leu Val Glu Arg Asn Trp 245 250 255

Ala Asn Val Phe Gly Thr Ser Asn Lys Met Ala Glu Leu Asn Tyr Lys 260 265 270 ❹ lie Cys Ala Arg Lys Met Leu Met lie Asp Arg Val Thr His lie Asp 275 280 285Ala Asn Val Phe Gly Thr Ser Asn Lys Met Ala Glu Leu Asn Tyr Lys 260 265 270 ❹ lie Cys Ala Arg Lys Met Leu Met lie Asp Arg Val Thr His lie Asp 275 280 285

His His Gly Gly Ala Tyr Gly Leu Gly Leu Leu Val Gly Glu Lys lie 290 295 300His His Gly Gly Ala Tyr Gly Leu Gly Leu Leu Val Gly Glu Lys lie 290 295 300

Leu Asp Arg Asn His Trp Tyr Phe Pro Cys His Phe Val Asn Asp Gin 305 310 315 320Leu Asp Arg Asn His Trp Tyr Phe Pro Cys His Phe Val Asn Asp Gin 305 310 315 320

Val Met Ala Gly Ser Leu Val Ser Asp Gly Cys Ser Gin Leu Leu Lys 325 330 335Val Met Ala Gly Ser Leu Val Ser Asp Gly Cys Ser Gin Leu Leu Lys 325 330 335

Leu Tyr Met lie Trp Leu Gly Leu His Leu Lys Met Glu Glu Phe Asp 340 345 350Leu Tyr Met lie Trp Leu Gly Leu His Leu Lys Met Glu Glu Phe Asp 340 345 350

Phe Leu Pro Val Ser Gly His Lys Asn Lys Val Arg Cys Arg Gly Gin 355 360 365 lie Ser Pro His Lys Gly Lys Leu Val Tyr Val Met Glu lie Lys Lys 370 375 380Phe Leu Pro Val Ser Gly His Lys Asn Lys Val Arg Cys Arg Gly Gin 355 360 365 lie Ser Pro His Lys Gly Lys Leu Val Tyr Val Met Glu lie Lys Lys 370 375 380

Met Gly Tyr Asp Gin Ala Ser Gly Ser Pro Tyr Ala lie Ala Asp Val 385 390 395 400Met Gly Tyr Asp Gin Ala Ser Gly Ser Pro Tyr Ala lie Ala Asp Val 385 390 395 400

Asp lie lie Asp Val Asn Glu Glu Leu Gly Gin Ser Phe Asp lie Asn 405 410 415Asp lie lie Asp Val Asn Glu Glu Leu Gly Gin Ser Phe Asp lie Asn 405 410 415

Asp Leu Ala Ser Tyr Gly Lys Gly Asp Leu Ser Lys Lys lie Val Val 420 425 430Asp Leu Ala Ser Tyr Gly Lys Gly Asp Leu Ser Lys Lys lie Val Val 420 425 430

Asp Phe Lys Gly lie Ala Leu Gin Leu Lys Gly Arg Ala Phe Ser Arg 435 440 445Asp Phe Lys Gly lie Ala Leu Gin Leu Lys Gly Arg Ala Phe Ser Arg 435 440 445

Met Ser Ser Ser Ser Ser Leu Asn Glu Gly Trp Gin Cys Val Pro Lys 450 455 460Met Ser Ser Ser Ser Ser Leu Asn Glu Gly Trp Gin Cys Val Pro Lys 450 455 460

Pro Ser Gin Arg Met Glu His Glu Gin Pro Pro Ala His Cys Leu Ala 465 470 475 480Pro Ser Gin Arg Met Glu His Glu Gin Pro Pro Ala His Cys Leu Ala 465 470 475 480

Ser Asp Pro Glu Ala Pro Ser Thr Val Thr Trp His Pro Met Ser Lys 117 201038734 485 490 495Ser Asp Pro Glu Ala Pro Ser Thr Val Thr Trp His Pro Met Ser Lys 117 201038734 485 490 495

Leu Pro Gly Asn Pro Thr Pro Phe Phe Ser Pro Ser Ser 500 505 〇 r p o r p r o y-i—_ T 5Leu Pro Gly Asn Pro Thr Pro Phe Phe Ser Pro Ser Ser 500 505 〇 r p o r p r o y-i-_ T 5

Arg Ala lie Cys Phe lie Pro Phe Pro Gly Asn Pro Leu Asp Asn Asn 515 520 525Arg Ala lie Cys Phe lie Pro Phe Pro Gly Asn Pro Leu Asp Asn Asn 515 520 525

Cys Lys Ala Gly Glu Met 530 e h p u 1—I G r e s t e M no s 4 A 5 r Y· T p r T n s A u e L 0 5 r 3 p 5Cys Lys Ala Gly Glu Met 530 e h p u 1—I G r e s t e M no s 4 A 5 r Y· T p r T n s A u e L 0 5 r 3 p 5

Met Cys Gly Lys Val Ser Asn Cys Leu Gly Pro Glu Phe Ala Arg Phe 545 550 555 560Met Cys Gly Lys Val Ser Asn Cys Leu Gly Pro Glu Phe Ala Arg Phe 545 550 555 560

r 5 h 6 T 5 n s A r e s s y' L p s Ar 5 h 6 T 5 n s A r e s s y' L p s A

Ser Arg Ser Pro Ala Phe Asp Leu Ala Leu Val 570 5Ί5Ser Arg Ser Pro Ala Phe Asp Leu Ala Leu Val 570 5Ί5

Thr Arg Val Val Glu Val Thr Asn Met Glu His Gly Lys 580 585 Θ o h 9 p 5Thr Arg Val Val Glu Val Thr Asn Met Glu His Gly Lys 580 585 Θ o h 9 p 5

n s A u e Ln s A u e L

Val Asp Cys Asn Pro Ser Lys Gly Thr Met Val Gly Glu Phe Asp Cys 595 600 605Val Asp Cys Asn Pro Ser Lys Gly Thr Met Val Gly Glu Phe Asp Cys 595 600 605

Pro Gin Asp Ala Trp Phe Phe Asp Gly Ser Cys Asn Asp Gly His Met 610 615 620 r e s r y T o 5 r 2 p 6 r h T n 5 13 G 6 u e L y 1 G e 1 I u I—I G t o e 3 M6Pro Gin Asp Ala Trp Phe Phe Asp Gly Ser Cys Asn Asp Gly His Met 610 615 620 r e s r y T o 5 r 2 p 6 r h T n 5 13 G 6 u e L y 1 G e 1 I u I—I G t o e 3 M6

Ser Gly Val Leu 640Ser Gly Val Leu 640

Thr Ser Val Leu Lys Ala Pro Leu Thr Met Asp Lys Asp Asp lie Leu 645 650 655Thr Ser Val Leu Lys Ala Pro Leu Thr Met Asp Lys Asp Asp lie Leu 645 650 655

Phe Arg Asn Leu Asp Ala Ser Ala Glu Met Val Arg Pro Asp Val Asp 660 665 670Phe Arg Asn Leu Asp Ala Ser Ala Glu Met Val Arg Pro Asp Val Asp 660 665 670

Val Arg Gly Lys Thr lie Arg Asn Val Thr Lys Cys Thr Gly Tyr Ala 675 680 685Val Arg Gly Lys Thr lie Arg Asn Val Thr Lys Cys Thr Gly Tyr Ala 675 680 685

Met Leu Gly Lys Met Gly lie His Arg Phe Thr Phe Glu Leu Ser Val 690 695 700Met Leu Gly Lys Met Gly lie His Arg Phe Thr Phe Glu Leu Ser Val 690 695 700

Asp Gly Val Val Phe Tyr Lys Gly Ser Thr Ser Phe Gly Trp Phe Thr 7 05 710 715 720Asp Gly Val Val Phe Tyr Lys Gly Ser Thr Ser Phe Gly Trp Phe Thr 7 05 710 715 720

Pro Glu Val Phe Ala Gin Gin Ala Gly Leu Asp Asn Gly Lys Lys Thr 725 730 735Pro Glu Val Phe Ala Gin Gin Ala Gly Leu Asp Asn Gly Lys Lys Thr 725 730 735

Glu Pro Trp Cys Lys Thr Asn Asn Thr Ser Val Arg Arg Val Glu lie 740 745 750Glu Pro Trp Cys Lys Thr Asn Asn Thr Ser Val Arg Arg Val Glu lie 740 745 750

Ala Ser Ala Lys Gly Lys Glu Gin Leu Thr Glu Lys Leu Pro Asp Ala 755 760 765Ala Ser Ala Lys Gly Lys Glu Gin Leu Thr Glu Lys Leu Pro Asp Ala 755 760 765

Thr Asn Ala Gin Val Leu Arg Arg Ser Glu Gin Cys Glu Tyr Leu Asp 770 775 780 118 201038734Thr Asn Ala Gin Val Leu Arg Arg Ser Glu Gin Cys Glu Tyr Leu Asp 770 775 780 118 201038734

Tyr Leu Asn lie Ala Pro Asp Ser Gly Leu His Gly Lys Gly Tyr Ala 785 790 795 800Tyr Leu Asn lie Ala Pro Asp Ser Gly Leu His Gly Lys Gly Tyr Ala 785 790 795 800

His Gly His Lys Asp Val Asn Pro Gin Asp Trp Phe Phe Ser Cys His 805 810 815His Gly His Lys Asp Val Asn Pro Gin Asp Trp Phe Phe Ser Cys His 805 810 815

Phe Trp Phe Asp Pro Val Met Pro Gly Ser Leu Gly lie Glu Ser Met 820 825 830Phe Trp Phe Asp Pro Val Met Pro Gly Ser Leu Gly lie Glu Ser Met 820 825 830

Phe Gin Leu lie Glu Ala Phe Ala Val Asp Gin Asn lie Pro Gly Glu 835 840 845Phe Gin Leu lie Glu Ala Phe Ala Val Asp Gin Asn lie Pro Gly Glu 835 840 845

Tyr Asn Val Ser Asn Pro Thr Phe Ala His Ala Pro Gly Lys Thr Ala 850 855 860Tyr Asn Val Ser Asn Pro Thr Phe Ala His Ala Pro Gly Lys Thr Ala 850 855 860

Trp Lys Tyr Arg Gly Gin Leu Thr Pro Lys Asn Arg Ala Met Asp Cys 865 870 875 880 o oTrp Lys Tyr Arg Gly Gin Leu Thr Pro Lys Asn Arg Ala Met Asp Cys 865 870 875 880 o o

Glu Val His lie Val Ser lie Thr Ala Ser Pro Glu Asn Gly Gly Tyr 885 890 895Glu Val His lie Val Ser lie Thr Ala Ser Pro Glu Asn Gly Gly Tyr 885 890 895

Val Asp lie Val Ala Asp Gly Ala Leu Trp Val Asp Gly Leu Arg Val 900 905 910Val Asp lie Val Ala Asp Gly Ala Leu Trp Val Asp Gly Leu Arg Val 900 905 910

Tyr Glu Ala Lys Glu Leu Arg Val Arg Val Val Ser Ala Lvs Pro Gin 915 920 925Tyr Glu Ala Lys Glu Leu Arg Val Arg Val Val Ser Ala Lvs Pro Gin 915 920 925

Ala lie Pro Asp Val Gin Gin Gin Pro Pro Ser Ala Lys Ala Asp Pro 930 935 940Ala lie Pro Asp Val Gin Gin Gin Pro Pro Ser Ala Lys Ala Asp Pro 930 935 940

Gly Lys Thr Gly Val Ala Leu Ser Pro Thr Gin Leu Arg Asp Val Leu 945 950 955 960Gly Lys Thr Gly Val Ala Leu Ser Pro Thr Gin Leu Arg Asp Val Leu 945 950 955 960

Leu Glu Val Asp Asn Pro Leu Tyr Leu Gly Val Glu Asn Ser Asn Leu 965 970 975Leu Glu Val Asp Asn Pro Leu Tyr Leu Gly Val Glu Asn Ser Asn Leu 965 970 975

Val Gin Phe Glu Ser Lys Pro Ala Thr Ser Ser Arg lie Val Ser lie 980 985 990Val Gin Phe Glu Ser Lys Pro Ala Thr Ser Ser Arg lie Val Ser lie 980 985 990

Lys Pro Cys Ser lie Ser Asp Leu Gly Asp Lys Ser Phe Met Glu Thr 995 1000 1005Lys Pro Cys Ser lie Ser Asp Leu Gly Asp Lys Ser Phe Met Glu Thr 995 1000 1005

Tyr Asn Val Ser Ala Pro Leu Tyr Thr Gly Ala Met Ala Lys Glv 1010 1015 1020 lie Ala Ser Ala Asp Leu Val lie Ala Ala Gly Lys Arg Lys lie 1025 1030 1035Tyr Asn Val Ser Ala Pro Leu Tyr Thr Gly Ala Met Ala Lys Glv 1010 1015 1020 lie Ala Ser Ala Asp Leu Val lie Ala Ala Gly Lys Arg Lys lie 1025 1030 1035

Leu Gly Ser Phe Gly Ala Gly Gly Leu Pro lie Ser lie Val Ara 1040 1045 1050Leu Gly Ser Phe Gly Ala Gly Gly Leu Pro lie Ser lie Val Ara 1040 1045 1050

Glu Ala Leu Glu Lys lie Gin Gin His Leu Pro His Gly Pro Tvr 1055 1060 1065Glu Ala Leu Glu Lys lie Gin Gin His Leu Pro His Gly Pro Tvr 1055 1060 1065

Ala Val Asn Leu lie His Ser Pro Phe Asp Ser Asn Leu Glu Lvs 1070 1075 1080 119 201038734Ala Val Asn Leu lie His Ser Pro Phe Asp Ser Asn Leu Glu Lvs 1070 1075 1080 119 201038734

Gly Asn Val Asp Leu Phe Leu Glu Met Gly Val Thr Val Val Glu 1085 1090 1095Gly Asn Val Asp Leu Phe Leu Glu Met Gly Val Thr Val Val Glu 1085 1090 1095

Cys Ser Ala Phe Met Glu Leu Thr Ala Gin Val Val Arg Tyr Arg 1100 1105 1110Cys Ser Ala Phe Met Glu Leu Thr Ala Gin Val Val Arg Tyr Arg 1100 1105 1110

Ala Ser Gly Leu Ser Lys Ser Ala Asp Gly Ser lie Arg lie Ala 1115 1120 1125Ala Ser Gly Leu Ser Lys Ser Ala Asp Gly Ser lie Arg lie Ala 1115 1120 1125

His Arg lie lie Gly Lys Val Ser Arg Thr Glu Leu Ala Glu Met 1130 1135 1140His Arg lie lie Gly Lys Val Ser Arg Thr Glu Leu Ala Glu Met 1130 1135 1140

Phe lie Arg Pro Ala Pro Gin His Leu Leu Gin Lys Leu Val Ala 1145 1150 1155Phe lie Arg Pro Ala Pro Gin His Leu Leu Gin Lys Leu Val Ala 1145 1150 1155

Ser Gly Glu Leu Thr Ala Glu Gin Ala Glu Leu Ala Thr Gin Val 1160 1165 1170Ser Gly Glu Leu Thr Ala Glu Gin Ala Glu Leu Ala Thr Gin Val 1160 1165 1170

Pro Val Ala Asp Asp 工le Ala Val Glu Ala Asp Ser Gly Gly His 1175 1180 1185Pro Val Ala Asp Asp Ale Val Glu Ala Asp Ser Gly Gly His 1175 1180 1185

Thr Asp Asn Arg Pro lie His Val lie Leu Pro Leu lie lie Asn 1190 1195 1200Thr Asp Asn Arg Pro lie His Val lie Leu Pro Leu lie lie Asn 1190 1195 1200

Leu Arg Asn Arg Leu His Lys Glu Leu Asp Tyr Pro Ser His Leu 1205 1210 1215Leu Arg Asn Arg Leu His Lys Glu Leu Asp Tyr Pro Ser His Leu 1205 1210 1215

Arg Val Arg Val Gly Ala Gly Gly Gly lie Gly Cys Pro Gin Ala 1220 1225 1230Arg Val Arg Val Gly Ala Gly Gly Gly lie Gly Cys Pro Gin Ala 1220 1225 1230

Ala Leu Ala Ala Phe Gin Met Gly Ala Ala Phe Leu lie Thr Gly 1235 1240 1245Ala Leu Ala Ala Phe Gin Met Gly Ala Ala Phe Leu lie Thr Gly 1235 1240 1245

Thr Val Asn Gin Leu Ala Arg Glu Ser Gly Thr Cys Asp Asn Val 1250 1255 1260Thr Val Asn Gin Leu Ala Arg Glu Ser Gly Thr Cys Asp Asn Val 1250 1255 1260

Arg Leu Gin Leu Ser Lys Ala Thr Tyr Ser Asp Val Cys Met Ala 1265 1270 1275Arg Leu Gin Leu Ser Lys Ala Thr Tyr Ser Asp Val Cys Met Ala 1265 1270 1275

Pro Ala Ala Asp Met Phe Asp Gin Gly Val Glu Leu Gin Val Leu 1280 1285 1290Pro Ala Ala Asp Met Phe Asp Gin Gly Val Glu Leu Gin Val Leu 1280 1285 1290

Lys Lys Gly Thr Leu Phe Pro Ser Arg Ala Lys Lys Leu Tyr Glu 1295 1300 1305Lys Lys Gly Thr Leu Phe Pro Ser Arg Ala Lys Lys Leu Tyr Glu 1295 1300 1305

Leu Phe Cys Lys Tyr Asp Ser Phe Glu Ala Met Pro Ala Glu Glu 1310 1315 1320Leu Phe Cys Lys Tyr Asp Ser Phe Glu Ala Met Pro Ala Glu Glu 1310 1315 1320

Leu Gin Arg Val Glu Lys Arg lie Phe Gin Lys Ser Leu Ala Glu 1325 1330 1335Leu Gin Arg Val Glu Lys Arg lie Phe Gin Lys Ser Leu Ala Glu 1325 1330 1335

Val Trp Gin Glu Thr Ser Asp Phe Tyr lie His Arg lie Lvs Asn 1340 1345 1350Val Trp Gin Glu Thr Ser Asp Phe Tyr lie His Arg lie Lvs Asn 1340 1345 1350

Pro Glu Lys lie Asn Arg Ala Ala Ser Asp Gly Lys Leu Lvs Met 1355 1360 1365 120 201038734Pro Glu Lys lie Asn Arg Ala Ala Ser Asp Gly Lys Leu Lvs Met 1355 1360 1365 120 201038734

Ser Leu Cys Phe Arg Trp Tyr Leu Gly Leu Ser Ser Phe Trp Ala 1370 1375 1380Ser Leu Cys Phe Arg Trp Tyr Leu Gly Leu Ser Ser Phe Trp Ala 1370 1375 1380

Asn Ser Gly Ala Gin Asp Arg Val Met Asp Tyr Gin He Trp Cys 1385 1390 1395Asn Ser Gly Ala Gin Asp Arg Val Met Asp Tyr Gin He Trp Cys 1385 1390 1395

Gly Pro Ala He Gly Ala Phe Asn Asp Phe Thr Lys Gly Thr Tyr 1400 1405 1410Gly Pro Ala He Gly Ala Phe Asn Asp Phe Thr Lys Gly Thr Tyr 1400 1405 1410

Leu Asp Val Thr Val Ala Lys Ser Tyr Pro Cys Val Ala Gin He 1415 1420 1425Leu Asp Val Thr Val Ala Lys Ser Tyr Pro Cys Val Ala Gin He 1415 1420 1425

Asn Leu Gin lie Leu Gin Gly Ala Ala Tyr Leu Lys Arq Leu Glv 1430 1435 1440Asn Leu Gin lie Leu Gin Gly Ala Ala Tyr Leu Lys Arq Leu Glv 1430 1435 1440

Val lie Arg Phe Asp Arg Met Leu Leu Gin Ala Val Asp lie Asp ❹ 〇 1445 1450 1455Val lie Arg Phe Asp Arg Met Leu Leu Gin Ala Val Asp lie Asp ❹ 〇 1445 1450 1455

Asp Pro Val Phe Thr Tyr Val Pro Thr Gin Pro Leu 1460 1465 1470 <210> 61 <211> 1503Asp Pro Val Phe Thr Tyr Val Pro Thr Gin Pro Leu 1460 1465 1470 <210> 61 <211> 1503

<212> PRT <213> 裂殖壺菌物種 <400> 61<212> PRT <213> Schizochytrium species <400> 61

Met Ala Leu Arg Val Lys Thr Asn Lys Lys Pro Cys Trp Glu Met Thr 15 10 15Met Ala Leu Arg Val Lys Thr Asn Lys Lys Pro Cys Trp Glu Met Thr 15 10 15

Lys Glu Glu Leu Thr Ser Gly Lys Thr Glu Val Phe Asn Tyr Glu Glu 20 25 30Lys Glu Glu Leu Thr Ser Gly Lys Thr Glu Val Phe Asn Tyr Glu Glu 20 25 30

Leu Leu Glu Phe Ala Glu Gly Asp lie Ala Lys Val Phe Gly Pro Glu 35 40 45Leu Leu Glu Phe Ala Glu Gly Asp lie Ala Lys Val Phe Gly Pro Glu 35 40 45

Phe Ala Val lie Asp Lys Tyr Pro Arg Arg Val Arg Leu Pro Ala Arg 50 55 60Phe Ala Val lie Asp Lys Tyr Pro Arg Arg Val Arg Leu Pro Ala Arg 50 55 60

Glu Tyr Leu Leu Val Thr Arg Val Thr Leu Met Asp Ala Glu Val Asn 65 70 75 80Glu Tyr Leu Leu Val Thr Arg Val Thr Leu Met Asp Ala Glu Val Asn 65 70 75 80

Asn Tyr Arg Val Gly Ala Arg Met Val Thr Glu Tyr Asp Leu Pro Val 85 90 95Asn Tyr Arg Val Gly Ala Arg Met Val Thr Glu Tyr Asp Leu Pro Val 85 90 95

Asn Gly Glu Leu Ser Glu Gly Gly Asp Cys Pro Trp Ma Val Leu ValAsn Gly Glu Leu Ser Glu Gly Gly Asp Cys Pro Trp Ma Val Leu Val

Glu Ser Gly Gin Cys Asp Leu Met Leu lie Ser Tyr Met Gly lie Asp 115 120 125Glu Ser Gly Gin Cys Asp Leu Met Leu lie Ser Tyr Met Gly lie Asp 115 120 125

Phe Gin Asn Gin Gly Asp Arg Val Tyr Arg Leu Leu Asn Thr Thr Leu 130 135 140Phe Gin Asn Gin Gly Asp Arg Val Tyr Arg Leu Leu Asn Thr Thr Leu 130 135 140

Thr Phe Tyr Gly Val Ala His Glu Gly Glu Thr Leu Glu Tyr Asp lie 145 150 155 160 121 201038734Thr Phe Tyr Gly Val Ala His Glu Gly Glu Thr Leu Glu Tyr Asp lie 145 150 155 160 121 201038734

Arg Val Thr Gly Phe Ala Lys Arg Leu Asp Gly Gly lie Ser Met Phe 165 170 175Arg Val Thr Gly Phe Ala Lys Arg Leu Asp Gly Gly lie Ser Met Phe 165 170 175

Phe Phe Glu Tyr Asp Cys Tyr Val Asn Gly Arg Leu Leu lie Glu Met 180 185 190Phe Phe Glu Tyr Asp Cys Tyr Val Asn Gly Arg Leu Leu lie Glu Met 180 185 190

Arg Asp Gly Cys Ala Gly Phe Phe Thr Asn Glu Glu Leu Asp Ala Gly 195 200 205Arg Asp Gly Cys Ala Gly Phe Phe Thr Asn Glu Glu Leu Asp Ala Gly 195 200 205

Lys Gly Val Val Phe Thr Arg Gly Asp Leu Ala Ala Arg Ala Lys lie 210 215 220Lys Gly Val Val Phe Thr Arg Gly Asp Leu Ala Ala Arg Ala Lys lie 210 215 220

Pro Lys Gin Asp Val Ser Pro Tyr Ala Val Ala Pro Cys Leu His Lys 225 230 235 240Pro Lys Gin Asp Val Ser Pro Tyr Ala Val Ala Pro Cys Leu His Lys 225 230 235 240

Thr Lys Leu Asn Glu Lys Glu Met Gin Thr Leu Val Asp Lys Asp Trp 245 250 255Thr Lys Leu Asn Glu Lys Glu Met Gin Thr Leu Val Asp Lys Asp Trp 245 250 255

Ala Ser Val Phe Gly Ser Lys Asn Gly Met Pro Glu lie Asn Tyr Lys 260 265 270Ala Ser Val Phe Gly Ser Lys Asn Gly Met Pro Glu lie Asn Tyr Lys 260 265 270

Leu Cys Ala Arg Lys Met Leu Met lie Asp Arg Val Thr Ser lie Asp 275 280 285Leu Cys Ala Arg Lys Met Leu Met lie Asp Arg Val Thr Ser lie Asp 275 280 285

His Lys Gly Gly Val Tyr Gly Leu Gly Gin Leu Val Gly Glu Lys lie 290 295 300His Lys Gly Gly Val Tyr Gly Leu Gly Gin Leu Val Gly Glu Lys lie 290 295 300

Leu Glu Arg Asp His Trp Tyr Phe Pro Cys His Phe Val Lys Asp Gin 305 310 315 320Leu Glu Arg Asp His Trp Tyr Phe Pro Cys His Phe Val Lys Asp Gin 305 310 315 320

Val Met Ala Gly Ser Leu Val Ser Asp Gly Cys Ser Gin Met Leu Lys 325 330 335Val Met Ala Gly Ser Leu Val Ser Asp Gly Cys Ser Gin Met Leu Lys 325 330 335

Met Tyr Met lie Trp Leu Gly Leu His Leu Thr Thr Gly Pro Phe Asp 340 345 350Met Tyr Met lie Trp Leu Gly Leu His Leu Thr Thr Gly Pro Phe Asp 340 345 350

Phe Arg Pro Val Asn Gly His Pro Asn Lys Val Arg Cys Arg Gly Gin 355 360 365 lie Ser Pro His Lys Gly Lys Leu Val Tyr Val Met Glu 工le Lys Glu 370 375 380Phe Arg Pro Val Asn Gly His Pro Asn Lys Val Arg Cys Arg Gly Gin 355 360 365 lie Ser Pro His Lys Gly Lys Leu Val Tyr Val Met Glu work le Lys Glu 370 375 380

Met Gly Phe Asp Glu Asp Asn Asp Pro Tyr Ala lie Ala Asp Val Asn 385 390 395 400 lie lie Asp Val Asp Phe Glu Lys Gly Gin Asp Phe Ser Leu Asp Arg 405 410 415 lie Ser Asp Tyr Gly Lys Gly Asp Leu Asn Lys Lys lie Val Val Asp 420 425 430Met Gly Phe Asp Glu Asp Asn Asp Pro Tyr Ala lie Ala Asp Val Asn 385 390 395 400 lie lie Asp Val Asp Phe Glu Lys Gly Gin Asp Phe Ser Leu Asp Arg 405 410 415 lie Ser Asp Tyr Gly Lys Gly Asp Leu Asn Lys Lys lie Val Val Asp 420 425 430

Phe Lys Gly lie Ala Leu Lys Met Gin Lys Arg Ser Thr Asn Lys Asn 435 440 445Phe Lys Gly lie Ala Leu Lys Met Gin Lys Arg Ser Thr Asn Lys Asn 435 440 445

Pro Ser Lys Val Gin Pro Val Phe Ala Asn Gly Ala Ala Thr Val Gly 122 201038734 450 455 460Pro Ser Lys Val Gin Pro Val Phe Ala Asn Gly Ala Ala Thr Val Gly 122 201038734 450 455 460

Pro Glu Ala Ser Lys Ala Ser Ser Gly Ala Ser Ala Ser Ala Ser Ala 465 470 475 480Pro Glu Ala Ser Lys Ala Ser Ser Gly Ala Ser Ala Ser Ala Ser Ala 465 470 475 480

Ala Pro Ala Lys Pro Ala Phe Ser Ala Asp Val Leu Ala Pro Lys Pro 485 490 495Ala Pro Ala Lys Pro Ala Phe Ser Ala Asp Val Leu Ala Pro Lys Pro 485 490 495

Val Ala Leu Pro Glu His lie Leu Lys Gly Asp Ala Leu Ala Pro Lys 500 505 510Val Ala Leu Pro Glu His lie Leu Lys Gly Asp Ala Leu Ala Pro Lys 500 505 510

Glu Met Ser Trp His Pro Met Ala Arg lie Pro Gly Asn Pro Thr Pro 515 520 525Glu Met Ser Trp His Pro Met Ala Arg lie Pro Gly Asn Pro Thr Pro 515 520 525

Ser Phe Ala Pro Ser Ala Tyr Lys Pro Arg Asn lie Ala Phe Thr Pro 530 535 540 ❹Ser Phe Ala Pro Ser Ala Tyr Lys Pro Arg Asn lie Ala Phe Thr Pro 530 535 540 ❹

Phe Pro Gly Asn Pro Asn Asp Asn Asp His Thr Pro Gly Lys Met Pro 545 550 555 560Phe Pro Gly Asn Pro Asn Asp Asn Asp His Thr Pro Gly Lys Met Pro 545 550 555 560

Leu Thr Trp Phe Asn Met Ala Glu Phe Met Ala Gly Lys Val Ser Met 565 570 575Leu Thr Trp Phe Asn Met Ala Glu Phe Met Ala Gly Lys Val Ser Met 565 570 575

Cys Leu Gly Pro Glu Phe Ala Lys Phe Asp Asp Ser Asn Thr Ser Arg 580 585 590Cys Leu Gly Pro Glu Phe Ala Lys Phe Asp Asp Ser Asn Thr Ser Arg 580 585 590

Ser Pro Ala Trp Asp Leu Ala Leu Val Thr Arg Ala Val Ser Val Ser 595 600 605Ser Pro Ala Trp Asp Leu Ala Leu Val Thr Arg Ala Val Ser Val Ser 595 600 605

Asp Leu Lys His Val Asn Tyr Arg Asn lie Asp Leu Asp Pro Ser Lys 610 615 620Asp Leu Lys His Val Asn Tyr Arg Asn lie Asp Leu Asp Pro Ser Lys 610 615 620

Gly Thr Met Val Gly Glu Phe Asp Cys Pro Ala Asp Ala Trp Phe Tyr 625 630 635 640Gly Thr Met Val Gly Glu Phe Asp Cys Pro Ala Asp Ala Trp Phe Tyr 625 630 635 640

Lys Gly Ala Cys Asn Asp Ala His Met Pro Tyr Ser lie Leu Met Glu 645 650 655 lie Ala Leu Gin Thr Ser Gly Val Leu Thr Ser Val Leu Lys Ala Pro 660 665 670Lys Gly Ala Cys Asn Asp Ala His Met Pro Tyr Ser lie Leu Met Glu 645 650 655 lie Ala Leu Gin Thr Ser Gly Val Leu Thr Ser Val Leu Lys Ala Pro 660 665 670

Leu Thr Met Glu Lys Asp Asp lie Leu Phe Arg Asn Leu Asp Ala Asn 675 680 685Leu Thr Met Glu Lys Asp Asp lie Leu Phe Arg Asn Leu Asp Ala Asn 675 680 685

Ala Glu Phe Val Arg Ala Asp Leu Asp Tyr Arg Gly Lys Thr lie Arg 690 695 700Ala Glu Phe Val Arg Ala Asp Leu Asp Tyr Arg Gly Lys Thr lie Arg 690 695 700

Asn Val Thr Lys Cys Thr Gly Tyr Ser Met Leu Gly Glu Met Gly Val 705 710 715 720Asn Val Thr Lys Cys Thr Gly Tyr Ser Met Leu Gly Glu Met Gly Val 705 710 715 720

His Arg Phe Thr Phe Glu Leu Tyr Val Asp Asp Val Leu Phe Tyr Lys 725 730 735His Arg Phe Thr Phe Glu Leu Tyr Val Asp Asp Val Leu Phe Tyr Lys 725 730 735

Gly Ser Thr Ser Phe Gly Trp Phe Val Pro Glu Val Phe Ala Ala Gin 740 745 750 123 201038734Gly Ser Thr Ser Phe Gly Trp Phe Val Pro Glu Val Phe Ala Ala Gin 740 745 750 123 201038734

Ala Gly Leu Asp Asn Gly Arg Lys Ser Glu Pro Trp Phe lie Glu Asn 755 760 765Ala Gly Leu Asp Asn Gly Arg Lys Ser Glu Pro Trp Phe lie Glu Asn 755 760 765

Lys Val Pro Ala Ser Gin Val Ser Ser Phe Asp Val Arg Pro Asn Gly 770 775 780Lys Val Pro Ala Ser Gin Val Ser Ser Phe Asp Val Arg Pro Asn Gly 770 775 780

Ser Gly Arg Thr Ala lie Phe Ala Asn Ala Pro Ser Gly Ala Gin Leu 785 790 795 800Ser Gly Arg Thr Ala lie Phe Ala Asn Ala Pro Ser Gly Ala Gin Leu 785 790 795 800

Asn Arg Arg Thr Asp Gin Gly Gin Tyr Leu Asp Ala Val Asp lie Val 805 810 815Asn Arg Arg Thr Asp Gin Gly Gin Tyr Leu Asp Ala Val Asp lie Val 805 810 815

Ser Gly Ser Gly Lys Lys Ser Leu Gly Tyr Ala His Gly Ser Lys Thr 820 825 830Ser Gly Ser Gly Lys Lys Ser Leu Gly Tyr Ala His Gly Ser Lys Thr 820 825 830

Val Asn Pro Asn Asp Trp Phe Phe Ser Cys His Phe Trp Phe Asp Ser 835 840 845Val Asn Pro Asn Asp Trp Phe Phe Ser Cys His Phe Trp Phe Asp Ser 835 840 845

Val Met Pro Gly Ser Leu Gly Val Glu Ser Met Phe Gin Leu Val Glu 850 855 860 fVal Met Pro Gly Ser Leu Gly Val Glu Ser Met Phe Gin Leu Val Glu 850 855 860 f

% I% I

Ala lie Ala Ala His Glu Asp Leu Ala Gly Lys Ala Arg His Cys Gin 865 870 875 880Ala lie Ala Ala His Glu Asp Leu Ala Gly Lys Ala Arg His Cys Gin 865 870 875 880

Pro His Leu Cys Ala Arg Pro Arg Ala Arg Ser Ser Trp Lys Tyr Arg 885 890 895Pro His Leu Cys Ala Arg Pro Arg Ala Arg Ser Ser Trp Lys Tyr Arg 885 890 895

Gly Gin Leu Thr Pro Lys Ser Lys Lys Met Asp Ser Glu Val His lie 900 905 910Gly Gin Leu Thr Pro Lys Ser Lys Lys Met Asp Ser Glu Val His lie 900 905 910

Val Ser Val Asp Ala His Asp Gly Val Val Asp Leu Val Ala Asp Gly 915 920 925Val Ser Val Asp Ala His Asp Gly Val Val Asp Leu Val Ala Asp Gly 915 920 925

Phe Leu Trp Ala Asp Ser Leu Arg Val Tyr Ser Val Ser Asn lie Arg 930 935 940Phe Leu Trp Ala Asp Ser Leu Arg Val Tyr Ser Val Ser Asn lie Arg 930 935 940

Val Arg lie Ala Ser Gly Glu Ala Pro Ala Ala Ala Ser Ser Ala Ala 945 950 955 960Val Arg lie Ala Ser Gly Glu Ala Pro Ala Ala Ala Ser Ser Ala Ala 945 950 955 960

Ser Val Gly Ser Ser Ala Ser Ser Val Glu Arg Thr Arg Ser Ser Pro 965 970 975Ser Val Gly Ser Ser Ala Ser Ser Val Glu Arg Thr Arg Ser Ser Pro 965 970 975

Ala Val Ala Ser Gly Pro Ala Gin Thr 工le Asp Leu Lys Gin Leu Lys 980 985 990Ala Val Ala Ser Gly Pro Ala Gin Thr work Le Asp Leu Lys Gin Leu Lys 980 985 990

Thr Glu Leu Leu Glu Leu Asp Ala Pro Leu Tyr Leu Ser Gin Asp Pro 995 1000 1005Thr Glu Leu Leu Glu Leu Asp Ala Pro Leu Tyr Leu Ser Gin Asp Pro 995 1000 1005

Thr Ser Gly Gin Leu Lys Lys His Thr Asp Val Ala Ser Gly Gin 1010 1015 1020Thr Ser Gly Gin Leu Lys Lys His Thr Asp Val Ala Ser Gly Gin 1010 1015 1020

Ala Thr lie Val Gin Pro Cys Thr Leu Gly Asp Leu Gly Asp Arg 1025 1030 1035Ala Thr lie Val Gin Pro Cys Thr Leu Gly Asp Leu Gly Asp Arg 1025 1030 1035

Ser Phe Met Glu Thr Tyr Gly Val Val Ala Pro Leu Tyr Thr Gly 1040 1045 1050 124 201038734Ser Phe Met Glu Thr Tyr Gly Val Val Ala Pro Leu Tyr Thr Gly 1040 1045 1050 124 201038734

Ala Met Ala Lys Gly lie Ala 1055 1060 Ser Ala Asp Leu Val lie Ala Ala 1065 Gly Lys Arg Lys lie Leu Gly 1070 1075 Ser Phe Gly Ala Gly Gly Leu Pro 1080 Met His His Val Arg Ala Ala 1085 1090 Leu Glu Lys lie Gin Ala Ala Leu 1095 Pro Gin Gly Pro Tyr Ala Val 1100 1105 Asn Leu lie His Ser Pro Phe Asp 1110 Ser Asn Leu Glu Lys Gly Asn 1115 1120 Val Asp Leu Phe Leu Glu Lys Gly 1125 Val Thr Val Val Glu Ala Ser 1130 1135 Ala Phe Met Thr Leu Thr Pro Gin 1140 O Val Val Arg Tyr Arg Ala Ala 1145 1150 Gly Leu Ser Arg Asn Ala Asp Gly 1155 Ser Val Asn lie Arg Asn Arg 1160 1165 lie lie Gly Lys Val Ser Arg Thr 1170 Glu Leu Ala Glu Met Phe lie • 1175 1180 Arg Pro Ala Pro Glu His Leu Leu 1185 - Glu Lys Leu lie Ala Ser Gly 1190 1195 Glu lie Thr Gin Glu Gin Ala Glu 1200 Leu Ala Arg Arg Val Pro Val 1205 1210 Ala Asp Asp 工le Ala Val Glu Ala 1215 Asp Ser Gly Gly His Thr Asp 1220 1225 Asn Arg Pro lie His Val lie Leu 1230 Pro Leu lie lie Asn Leu Arg ❹ 1235 1240 Asn Arg Leu His Arg Glu Cys Gly 1245 Tyr Pro Ala His Leu Arg Val 1250 1255 Arg Val Gly Ala Gly Gly Gly Val 1260 Gly Cys Pro Gin Ala Ala Ala 1265 1270 Ala Ala Leu Thr Met Gly Ala Ala 1275 Phe lie Val Thr Gly Thr Val 1280 1285 Asn Gin Val Ala Lys Gin Ser Gly 1290 Thr Cys Asp Asn Val Arg Lys 1295 1300 Gin Leu Ser Gin Ala Thr Tyr Ser 1305 Asp lie Cys Met Ala Pro Ala 1310 1315 Ala Asp Met Phe Glu Glu Gly Val 1320 Lys Leu Gin Val Leu Lys Lys 1325 1330 Gly Thr Met Phe Pro Ser Arg Ala 1335 125 201038734Ala Met Ala Lys Gly lie Ala 1055 1060 Ser Ala Asp Leu Val lie Ala Ala 1065 Gly Lys Arg Lys lie Leu Gly 1070 1075 Ser Phe Gly Ala Gly Gly Leu Pro 1080 Met His His Val Arg Ala Ala 1085 1090 Leu Glu Lys lie Gin Ala Ala Leu 1095 Pro Gin Gly Pro Tyr Ala Val 1100 1105 Asn Leu lie His Ser Pro Phe Asp 1110 Ser Asn Leu Glu Lys Gly Asn 1115 1120 Val Asp Leu Phe Leu Glu Lys Gly 1125 Val Thr Val Val Glu Ala Ser 1130 1135 Ala Phe Met Thr Leu Thr Pro Gin 1140 O Val Val Arg Tyr Arg Ala Ala 1145 1150 Gly Leu Ser Arg Asn Ala Asp Gly 1155 Ser Val Asn lie Arg Asn Arg 1160 1165 lie lie Gly Lys Val Ser Arg Thr 1170 Glu Leu Ala Glu Met Phe lie • 1175 1180 Arg Pro Ala Pro Glu His Leu Leu 1185 - Glu Lys Leu lie Ala Ser Gly 1190 1195 Glu lie Thr Gin Glu Gin Ala Glu 1200 Leu Ala Arg Arg Val Pro Val 1205 1210 Ala Asp Asp Ale Val Glu Ala 1215 Asp Ser Gly Gly His Thr Asp 1220 1225 Asn Arg Pro lie His Val lie Leu 1230 Pro Leu lie lie Asn Leu Arg ❹ 1235 1240 Asn Arg Leu His Arg Glu Cys Gly 1245 Tyr Pro Ala His Leu Arg Val 1250 1255 Arg Val Gly Ala Gly Gly Gly Val 1260 Gly Cys Pro Gin Ala Ala Ala 1265 1270 Ala Ala Leu Thr Met Gly Ala Ala 1275 Phe lie Val Thr Gly Thr Val 1280 1285 Asn Gin Val Ala Lys Gin Ser Gly 1290 Thr Cys Asp Asn Val Arg Lys 1295 1300 Gin Leu Ser Gin Ala Thr Tyr Ser 1305 Asp lie Cys Met Ala Pro Ala 1310 1315 Ala Asp Met Phe Glu Glu Gly Val 1320 Lys Leu Gin Val Leu Lys Lys 1325 1330 Gly Thr Met Phe Pro Ser Arg Ala 1335 125 201038734

Asn Lys Leu Tyr Glu Leu Phe Cys Lys Tyr 1340 1345Asn Lys Leu Tyr Glu Leu Phe Cys Lys Tyr 1340 1345

Asp Ser Phe Asp Ser 1350Asp Ser Phe Asp Ser 1350

Met Pro Pro Ala Glu Leu Glu Arg lie Glu 1355 1360Met Pro Pro Ala Glu Leu Glu Arg lie Glu 1355 1360

Lys Arg lie Phe Lys 1365Lys Arg lie Phe Lys 1365

Arg Ala Leu Gin Glu Val Trp Glu Glu Thr 1370 1375Arg Ala Leu Gin Glu Val Trp Glu Glu Thr 1370 1375

Lys Asp Phe Tyr lie 1380Lys Asp Phe Tyr lie 1380

Asn Gly Leu Lys Asn Pro Glu Lys lie Gin 1385 1390Asn Gly Leu Lys Asn Pro Glu Lys lie Gin 1385 1390

Arg Ala Glu His Asp 1395Arg Ala Glu His Asp 1395

Pro Lys Leu Lys Met Ser Leu Cys Phe Arg 1400 1405Pro Lys Leu Lys Met Ser Leu Cys Phe Arg 1400 1405

Trp Tyr Leu Gly Leu 1410Trp Tyr Leu Gly Leu 1410

Ala Ser Arg Trp Ala Asn Met Gly Ala Pro 1415 1420Ala Ser Arg Trp Ala Asn Met Gly Ala Pro 1415 1420

Asp Arg Val Met Asp 1425Asp Arg Val Met Asp 1425

Tyr Gin Val Trp Cys Gly Pro Ala lie Gly 1430 1435Tyr Gin Val Trp Cys Gly Pro Ala lie Gly 1430 1435

Ala Phe Asn Asp Phe 1440 lie Lys Gly Thr Tyr Leu Asp Pro Ala Val 1445 1450Ala Phe Asn Asp Phe 1440 lie Lys Gly Thr Tyr Leu Asp Pro Ala Val 1445 1450

Ser Asn Glu Tyr Pro 1455Ser Asn Glu Tyr Pro 1455

Cys Val Val Gin 工le Asn Leu Gin lie Leu 1460 1465Cys Val Val Gin work le Asn Leu Gin lie Leu 1460 1465

Arg Gly Ala Cys Tyr 1470Arg Gly Ala Cys Tyr 1470

Leu Arg Arg Leu Asn Ala Leu Arg Asn Asp 1475 1480Leu Arg Arg Leu Asn Ala Leu Arg Asn Asp 1475 1480

Pro Arg lie Asp Leu 1485Pro Arg lie Asp Leu 1485

Glu Thr Glu Asp Ala Ala Phe Val Tyr Glu 1490 1495Glu Thr Glu Asp Ala Ala Phe Val Tyr Glu 1490 1495

Pro Thr Asn Ala Leu 1500 <210> 62 <211> 27 <212> DNA <213> 人工序列 <220> <223> 合成引子prDS173Pro Thr Asn Ala Leu 1500 <210> 62 <211> 27 <212> DNA <213> Artificial sequence <220><223> Synthetic primer prDS173

> > > > 0 12 3 2 2 2 2 2 2 2 2 < < < < 其他特徵 (19)..(19) n 為 a、c、g，或 t <220> <221> 其他特徵 <222> (22)..(22) <223〉 n 為 a、c、g，或 t <400> 62 gatctactgc aagcgcggng gnttyat 27 <210> <211〉 <212> <213> 列 序<:工 31N V 6 2 D / 126 <220〉 201038734 <223〉合成引子prDS174 <220> <221> 其他特徵 <222> (16)..(16) <223> η 為 a、c、g，或 t <220> <221> 其他特徵 <222> (19) .. (19) <223> η 為 a、c、g，或 t <400> 63 ggcgcaggcg gcrtcnacna c <210> 64 <211> 23 <212> DNA <213> 人工序列 <220> <223> 合成引子JGM190 <220> <221> 其他特徵 <222> (15) . . (15) <223> n 為 a、c、g，或 t <400> 64 caytggtayt tyccntgyca ytt <210> 65 <211> 17 <212> DNA <213> 人工序列 <220> <223> 合成引子BLR242 <220> <221> 其他特徵 <222> (3)..(3) <223> n 為 a、c、g，或 t > > > > 0 12 3 2 2 2 2 2 2 2 2 < V < < 其他特徵 (9)..(9) η 為 a、c、g，或 t <220> <221> 其他特徵 <222> (12)..(12) <223> η 為 a、c、g，或 t <4〇〇> 65 ccnggcatna cnggrtc <210> 66 <211> 27 <212> DNA <213> 人工序列 <220> <223> 合成引子CAX055 <400> 66 qtcatgattg aacaagatgg attgcac 27 201038734 7 7Ny 6 2 D Λ 列 序 JA工 <210〉 <211> <212〉 <213〉 <220> <223> 合成引子CAX056 <400> 67 ccacgtgtca gaagaactcg tcaagaa <210> 68 <211> 8976 <212> DNA <213〉破囊壺菌物種 <400> 68 atggaagatc aaagaattgc tattgttgga ttatctgcga ttttaccaag tggtgaaaat 60 gttagagaat cttgggaagc aatacgtgat ggtttgaatt gtttaagtga tttacctgcg 120 gatcgtgttg atgttactgc gtattataat ccaacaaaag gtgtaaagga taaaatttat 180 tgtaaacgtg gtgggtttat tcctgaatat gaatttgatt ctagagaatt tggacttaat 240 atgttacaaa tggaagattc tgatgctaat caaacgttaa ctttattaaa ggttaaagaa 300 gcattagatg atgctaatat acctgcattt actaatgaga aaaaaaatat tggttgtgtt 360 cttggtattg gtggtggtca aaaagcatct catgaatttt attcaagact taattatgtt 420 gttgtggata aagttttaag aaaaatggga ttacctgatg aggatgttga aactgctgtt 480 gaaaagttta aagctaattt tcctgaatgg agattagatt cctttcctgg ttttcttggt 540 aatgttactg ctggccgttg tactaataca ttcaatatgg aaggtatgaa ttgtgttgta 600 gatgctgctt gtgctagttc tttaattgct attaaagttg ctattgatga attattacat 660 ggtgattgtg atgcaatgat tgctggtgca acttgtactg ataacgctct tggtatgtat 720 atggcatttt caaaaacacc tgttttttca actgatcaaa gttgtcttgc atatgatgaa 780 aaaacaaaag gtatgcttat tggtgaaggt tcagctatgt ttgttttaaa acgttatgct 840 gacgcagtga gagatggtga tactgtacat gctgttatac gttcatgttc atcatcatct 900 gacggtaaag catctggtat ttatacacca actatttctg gtcaagaaga agctattctt 960 agagcatatc gtagagctgg tgtatcacca aatactatta ctttagttga aggacatggt 1020 actggtacac cagtgggtga taaaattgaa ttaacagctt tacgcaatgt atttgataaa 1080 gcatatggtc ctggtcataa ggaagaagtt gctgttggaa gtattaaaag tcaaattggt 1140 catttgaaag ctgttgctgg ttgtgctggt cttgtgaaat tggttatggc attgaaacat 1200 aaaacactac ctcaaagtat taatgttgaa aatccaccta atttagtgga tggtactgtc 1260 attagtgata ctactttata tattaataca atgaatcgtc catggattac taagcctggt 1320 gttccaagaa gagctggtat atctagtttc ggatttggtg gtgcaaatta tcatgctgtt 1380 ttagaagaat ttgagccgga acaaactaaa ccatatagat tgaatgtatc tgcacaacca 1440 atgcttcttc atgcggtaaa tgcaaattca ttacaaaagc tatgtgaaga tcaattaasa 15〇0 cttttgaaag aatcaagaga aaaatgtgtc aacaccaaaa acactgatta tgttgcgttt 1560 tcaaaatttc aagattcttt taaattgaaa ggttctgttc catcacaaca tgctagagtt 1620 ggttttgcat caaaatctat tgaagatact atttctattt tatctgctat cgttaataga 1680 128 201038734>>>> 0 12 3 2 2 2 2 2 2 2 2 <<<< Other characteristics (19).. (19) n is a, c, g, or t < 220 >;<221> Other Features <222> (22)..(22) <223> n is a, c, g, or t <400> 62 gatctactgc aagcgcggng gnttyat 27 <210><211><212><213> Column order <: work 31N V 6 2 D / 126 < 220> 201038734 <223> Synthesis primer prDS174 <220><221> Other features <222> (16) ..(16) <223> η is a, c, g, or t <220><221> Other features <222> (19) .. (19) <223> η is a, c , g, or t <400> 63 ggcgcaggcg gcrtcnacna c <210> 64 <211> 23 <212> DNA <213> Artificial sequence <220><223> Synthetic primer JGM190 <220>;221> Other Features <222> (15) . . . (15) <223> n is a, c, g, or t <400> 64 caytggtayt tyccntgyca ytt <210> 65 <211> 17 <;212> DNA <213> Artificial Sequence <220><223> Synthesis Primer BLR242 <220><221> Other characteristics <222> (3)..(3) <223> n is a, c, g, or t >>>> 0 12 3 2 2 2 2 2 2 2 2 < V << Other characteristics (9)..(9) η is a, c, g, or t <220><221> Other characteristics <222> (12)..(12) <223> η is a, c, g, or t <4〇〇> 65 ccnggcatna cnggrtc <210> 66 <211> 27 <212> DNA <213> Artificial sequence <220>;223> Synthetic primer CAX055 <400> 66 qtcatgattg aacaagatgg attgcac 27 201038734 7 7Ny 6 2 D Λ Sequence JA <210> <211><212><213><220><223> Synthetic primer CAX056 <400> 67 ccacgtgtca gaagaactcg tcaagaa <210> 68 <211> 8976 <212> DNA <213> Thraustochytrium species <400> 68 atggaagatc aaagaattgc tattgttgga ttatctgcga ttttaccaag tggtgaaaat 60 gttagagaat cttgggaagc aatacgtgat Ggtttgaatt gtttaagtga tttacctgcg 120 gatcgtgttg atgttactgc gtattataat ccaacaaaag gtgtaaagga taaaatttat 180 tgtaaacgtg gtgggtttat tcctgaatat gaatttg att ctagagaatt tggacttaat 240 atgttacaaa tggaagattc tgatgctaat caaacgttaa ctttattaaa ggttaaagaa 300 gcattagatg atgctaatat acctgcattt actaatgaga aaaaaaatat tggttgtgtt 360 cttggtattg gtggtggtca aaaagcatct catgaatttt attcaagact taattatgtt 420 gttgtggata aagttttaag aaaaatggga ttacctgatg aggatgttga aactgctgtt 480 gaaaagttta aagctaattt tcctgaatgg agattagatt cctttcctgg ttttcttggt 540 aatgttactg ctggccgttg tactaataca ttcaatatgg aaggtatgaa ttgtgttgta 600 gatgctgctt gtgctagttc tttaattgct attaaagttg ctattgatga attattacat 660 ggtgattgtg atgcaatgat tgctggtgca acttgtactg ataacgctct tggtatgtat 720 atggcatttt caaaaacacc tgttttttca actgatcaaa gttgtcttgc atatgatgaa 780 aaaacaaaag gtatgcttat tggtgaaggt tcagctatgt ttgttttaaa atcatcatct acgttatgct 840 gacgcagtga gagatggtga tactgtacat gctgttatac gttcatgttc actggtacac cagtgggtga taaaattgaa 900 gacggtaaag catctggtat ttatacacca actatttctg gtcaagaaga agctattctt 960 agagcatatc gtagagctgg tgtatcacca aatactatta ctttagttga aggacatggt 1020 ttaacagctt tacgcaatgt Attt gataaa 1080 gcatatggtc ctggtcataa ggaagaagtt gctgttggaa gtattaaaag tcaaattggt 1140 catttgaaag ctgttgctgg ttgtgctggt cttgtgaaat tggttatggc attgaaacat 1200 aaaacactac ctcaaagtat taatgttgaa aatccaccta atttagtgga tggtactgtc 1260 attagtgata ctactttata tattaataca atgaatcgtc catggattac taagcctggt 1320 gttccaagaa gagctggtat atctagtttc ggatttggtg gtgcaaatta tcatgctgtt 1380 ttagaagaat ttgagccgga acaaactaaa ccatatagat tgaatgtatc tgcacaacca 1440 atgcttcttc atgcggtaaa tgcaaattca ttacaaaagc tatgtgaaga tcaattaasa 〇 c c c c c c c c c 60 60 60 60 60 60 60 60 60 60 60 60 60 60 60 60

Ο tttcaaaaag atattacaac tctactgcat tgattaatga caatataccc atatgtttaa aatgatatgg agaaagcaca caagttatgt ttcctcgtaa tctaagactg aatattctca ttccgtgatg ctggtttcgc gcattgtatg cagctggatt gcaatggcta tgagagatgc ggtccaaatg cttcttcaat tctccatctc aaactgttat ttgaaaactc aaggtttccg catatggaaa atgctgaaaa ccaactggtt cttctccaaa aaaactgctt tgtcaagaca atgtacgcgg ctggagctcg ttggtcaatg aaatttttcc agtgctaaag atagtgacat gtagctctta ccgattttga ccacgtaaga agactacttt caggagcgtg aacgaatcat aacactaata ctggtgagtt gttgtaagag ttcaagctct gaattacaaa aagctcaagc aaacataagg caattttatt tattcttcat tttcgaaagg cctgttcaag cggctgctcc aaagcggaac aagttgtatt attgaattgg atatggaatt attctttctg aagttcaagc agaactcgta ctgttggtga ccagctgctc ctgttcaagt gctcctgttg attctggttt tcgaagactg gttatgagac ggtattgatt ctatcaagag gaagctaaag atgtagatgc atgaaagccg aaattgctgg caagtagttg ctcctgttca aactagttgg gctttaccaa caataaaagt gtagctgctt tgatgttgca atgcaatggc ggaagaagtt atcaatgata accatatgca agagaatcac aacaacaact gtcgctagtt tcctgctttt gttgctggtc gattgatcgc gaagatttat accaaaaaaa tctgctgatg taagctttca gctcctgaag taccggtgca aattctggtg tgtggttcat ttggcatgtg gcaatttcaa aaagctcttt aattttcagc aatgtaactg tatgactagt tctgtacaat tgtctttatt gaatttggac tggtgataca agcgttttaa tcaattgcgt caagctgcag taaatgggaa ctcaaagatc gactttgtct gcagcaactt gaatgatggg cgaactgttt ggaga33ttg 彐Bgaagc&at tgcaactcaa gcttcagctg aacaaaatct agtaatgcag ggcaatgtta gaagaacttg tcaagttgca agtccagcta tgtgcaggta tctgcttctg ggaagtattg gcatcgaaga ggaaactgaa cttggtattg tcaattgaat gttgaagcta agtgattgat gcaatgaaag tgcagctcct actcaagtag gttagcaaaa gcggaacaag tgagttgatt gaattggata agtagaaatt ctttctgaag tcttagtaga actcgtactg tggtcaacca gctgctcctg agcatctgct cctgttgatt aagaaggtgc tatttttaga tattttctgg acaaggcgca cacaatttcg tttatgtgta aaagtgtgaa acgtatcagt ctttagacaa taaagaaatc cagtaggttt atttgaaatt attctttagg tgaatttagt tcaagttggt atgtaatcgt gagcaatggc tgctgttatt tatgggttgc taacaataac tacaagctga aacaagtaaa atggggcatt tcattcgcct cagcagttaa gtttaataaa gtggtgtatt tacggatcca ttcttactca aattaagaat caaaacaagt actttccaaa ctgtttcggt gaatccagct ttcaaatggc cgttgctggt ctacccgtat gaaggaattc atgtctccaa gaaaactcta catgtgttca acgtattgaa tgcaagataa agaaaatgag atttgcaaaa taccaaagca catctgatgc ggtggtggca aaaccggcaa ggctgtagat ccgttcgtgt cgtttcagct ttgattctgg tttgttggca ctggttatga gactgagttg attctatcaa gagagtagaa aagatgtaga tgctcttagt ccgaaattgc tggtggtcaa ttgctcctgt tcaagcatct ttgtattgga agtattggca tggaattgga aaccgaactt ttcaagctca attgagtgtt ttggtgaagt gattgatgca ttcaagttgc agctcctact ctggtttgtt agcaaaagcg 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 129 201038734 gaacaagttg tattggaagt attggcatcg aagactggtt atgagactga gttgattgaa 4020 ttggatatgg aattggaaac cgaacttggt attgattcta tcaagagagt agaaattctt 4080 tctgaagttc aagctcaatt gagtgttgaa gctaaagatg tagatgctct tagtagaact 4140 cgtactgttg gtgaagtgat tgatgcaatg aaagctgaaa tttctggtgg tcagccagct 4200 gctcctgttc aagttgcagc tcctactcaa atagttgctc ctgttcaagt atccgctcct 4260 gttgattctg gtttgttagc aaaggcggaa caagtagtat tggaagtatt ggcatccaag 4320 actggttatg agactgagtt gattgaattg gatatggaat tggaaactga acttggtatt 4380 gattctatca agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct 4440 aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 4500 gctgaaattt ctggtggtca accaactgct cctgttcaag ttgcagctcc tactcaaata 4560 gttgctcctg ttcaagtatc tgctcctgtt gattctggtt tgttagcaaa ggcggaacaa 4620 gttgtattgg aagtattggc atcgaagact ggttatgaga ctgagttgat tgaattggat 4680 atggaattgg aaaccgaact tggtattgat tctatcaaga gagtagaaat tctttctgaa 4740 gttcaagctc aattgagtgt tgaagctaaa gatgtagatg ctcttagtag aactcgtact 4800 gttggtgaag tgattgatgc aatgaaagcc gaaatttctg gtggtcagcc agctgctcct 4860 gttcaagttg cagctcctac tcaaatagtt gctcctgttc aagcatctgc tcctgttgat 4920 tctggtttgt tggcaaaagc ggaacaagtt gtattggaag tgttagcatc caagactggt 4980 tatgaaactg agttgattga attagatatg gaattggaaa ccgaacttgg tattgattct 5040 atcaagagag tagaaattct ttctgaagtt caagctcaat tgagtgttga agctaaagat 5100 gtagatgctc ttagtagaac tcgtactgtt ggtgaagtga ttgatgcaat gaaagctgaa 5160 atttctggtg gtcaaccagc tgctcctgtt caagttgcag ctcctactca aatagttgct 5220 cctgttcaag tatctgctcc tgttgattct ggtttgttag caaaggcgga acaagttgta 5280 ttggaagtat tggcatctaa gactggttat gagactgagt tgattgaatt ggatatggaa 5340 ttggaaactg aacttggtat tgattctatc aagagagtag aaattctttc tgaagttcaa 5400 gctcaattga atgttgaagc taaagatgta gatgctctta gtagaactcg tactgttggt 5460 gaagtgattg atgcaatgaa agccgaaatt gctggtggtc aaccagctgc tcctgttcaa 5520 gttgcagctc ctgctccagt agttgctcct gttcaagtat ctactcctgt tgattctggt 5580 ttgttggcaa aagcggaaca agttgtattg gaagtgttag catgcaagac tggttatgaa 5640 actgagttga ttgaattgga tatggaattg gaaactgaac ttggtattga ttctatcaag 5700 agagtagaaa ttctttctga agttcaagct caattgagtg ttgaagctaa agatgtagat 5760 gctcttagta gaactcgtac tgttggtgaa gtgattgatg caatgaaagc cgaaatttct 5820 ggtggtcaac caactgctcc tgttcaagtt gcagctccta ctcaagtagt tgctcctgtt 5880 aaagtatcta ctcctgttga ttctggtttg ttagcaaagg cggaacaagt agtattggaa 5940 gtattggcat ctaagactgg ttatgaaact gagttgattg aattagatat ggaattggaa 6000 actgaacttg gtattgattc tatcaagaga gtagaaattc tttctgaagt tcaagctcaa 6060 ttgaatgtgg aagctaaaga tgtggatgct cttagtagaa ctcgtactgt tggtgaagtg 6120 attgatgcaa tgaaagccga aattgctggt gatcaacctg ctccagctgt agttccagtt 6180Ο tttcaaaaag atattacaac tctactgcat tgattaatga atatgtttaa caatataccc gttgtaagag ttcaagctct gaattacaaa aatgatatgg agaaagcaca caagttatgt ttcctcgtaa tctaagactg aatattctca ttccgtgatg ctggtttcgc gcattgtatg cagctggatt gcaatggcta tgagagatgc ggtccaaatg cttcttcaat tctccatctc aaactgttat ttgaaaactc aaggtttccg catatggaaa atgctgaaaa ccaactggtt cttctccaaa aaaactgctt tgtcaagaca atgtacgcgg ctggagctcg ttggtcaatg aaatttttcc agtgctaaag atagtgacat gtagctctta ccgattttga ccacgtaaga agactacttt caggagcgtg aacgaatcat aacactaata ctggtgagtt aagctcaagc aaacataagg caattttatt tattcttcat tttcgaaagg cctgttcaag cggctgctcc aaagcggaac aagttgtatt attgaattgg atatggaatt attctttctg aagttcaagc agaactcgta ctgttggtga ccagctgctc ctgttcaagt gctcctgttg attctggttt tcgaagactg gttatgagac ggtattgatt ctatcaagag gaagctaaag atgtagatgc atgaaagccg aaattgctgg caagtagttg ctcctgttca aactagttgg gctttaccaa caataaaagt gtagctgctt tgatgttgca atgcaatggc ggaagaagtt atcaatgata accatatgca agagaatcac aacaacaact gtcgctagtt tcctgctttt gttgctggtc gattgat cgc gaagatttat accaaaaaaa tctgctgatg taagctttca gctcctgaag taccggtgca aattctggtg tgtggttcat ttggcatgtg gcaatttcaa aaagctcttt aattttcagc aatgtaactg tatgactagt tctgtacaat tgtctttatt gaatttggac tggtgataca agcgttttaa tcaattgcgt caagctgcag taaatgggaa ctcaaagatc gactttgtct gcagcaactt gaatgatggg cgaactgttt ggaga33ttg Ji Bgaagc & at tgcaactcaa gcttcagctg aacaaaatct agtaatgcag ggcaatgtta gaagaacttg tcaagttgca agtccagcta tgtgcaggta tctgcttctg ggaagtattg gcatcgaaga ggaaactgaa cttggtattg tcaattgaat gttgaagcta agtgattgat gcaatgaaag tgcagctcct actcaagtag gttagcaaaa gcggaacaag tgagttgatt gaattggata agtagaaatt ctttctgaag tcttagtaga actcgtactg tggtcaacca gctgctcctg agcatctgct cctgttgatt aagaaggtgc tatttttaga tattttctgg acaaggcgca cacaatttcg tttatgtgta aaagtgtgaa acgtatcagt ctttagacaa taaagaaatc cagtaggttt atttgaaatt attctttagg tgaatttagt tcaagttggt atgtaatcgt gagcaatggc tgctgttatt tatgggttgc taacaataac tacaagctga aacaagtaaa atggggcatt tcattcgcct cagcagttaa gtttaataaa gtggtgtatt tacggatcca ttcttactca aattaagaat caaaacaagt actttccaaa ctgtttcggt gaatccagct ttcaaatggc cgttgctggt ctacccgtat gaaggaattc atgtctccaa gaaaactcta catgtgttca acgtattgaa tgcaagataa agaaaatgag atttgcaaaa taccaaagca catctgatgc ggtggtggca aaaccggcaa ggctgtagat ccgttcgtgt cgtttcagct ttgattctgg tttgttggca ctggttatga gactgagttg attctatcaa gagagtagaa aagatgtaga tgctcttagt ccgaaattgc tggtggtcaa ttgctcctgt tcaagcatct ttgtattgga agtattggca tggaattgga aaccgaactt ttcaagctca attgagtgtt ttggtgaagt gattgatgca ttcaagttgc agctcctact ctggtttgtt agcaaaagcg 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 129 201038734 gaacaagttg tattggaagt attggcatcg aagactggtt atgagactga gttgattgaa 4020 ttggatatgg aattggaaac cgaacttggt attgattcta tcaagagagt agaaattctt 4080 tctgaagttc aagctcaatt gagtgttgaa gctaaagatg tagatgctct tagtagaact 4140 cgtactgttg gtgaagtgat tgatgcaatg aaagctgaaa tttctggtgg tcagccag ct 4200 gctcctgttc aagttgcagc tcctactcaa atagttgctc ctgttcaagt atccgctcct 4260 gttgattctg gtttgttagc aaaggcggaa caagtagtat tggaagtatt ggcatccaag 4320 actggttatg agactgagtt gattgaattg gatatggaat tggaaactga acttggtatt 4380 gattctatca agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct 4440 aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 4500 gctgaaattt ctggtggtca accaactgct cctgttcaag ttgcagctcc tactcaaata 4560 gttgctcctg ttcaagtatc tgctcctgtt gattctggtt tgttagcaaa ggcggaacaa 4620 gttgtattgg aagtattggc atcgaagact ggttatgaga ctgagttgat tgaattggat 4680 atggaattgg aaaccgaact tggtattgat tctatcaaga gagtagaaat tctttctgaa 4740 gttcaagctc aattgagtgt tgaagctaaa gatgtagatg ctcttagtag aactcgtact 4800 gttggtgaag tgattgatgc aatgaaagcc gaaatttctg gtggtcagcc agctgctcct 4860 gttcaagttg cagctcctac tcaaatagtt gctcctgttc aagcatctgc tcctgttgat 4920 tctggtttgt tggcaaaagc ggaacaagtt gtattggaag tgttagcatc caagactggt 4980 tatgaaactg agttgattga attagatatg gaattggaaa ccgaacttgg tattgattct 5040 atcaagagag tagaaattct ttctgaagtt caagctcaat tgagtgttga agctaaagat 5100 gtagatgctc ttagtagaac tcgtactgtt ggtgaagtga ttgatgcaat gaaagctgaa 5160 atttctggtg gtcaaccagc tgctcctgtt caagttgcag ctcctactca aatagttgct 5220 cctgttcaag tatctgctcc tgttgattct ggtttgttag caaaggcgga acaagttgta 5280 ttggaagtat tggcatctaa gactggttat gagactgagt tgattgaatt ggatatggaa 5340 ttggaaactg aacttggtat tgattctatc aagagagtag aaattctttc tgaagttcaa 5400 gctcaattga atgttgaagc taaagatgta gatgctctta gtagaactcg tactgttggt 5460 gaagtgattg atgcaatgaa agccgaaatt gctggtggtc aaccagctgc tcctgttcaa 5520 gttgcagctc ctgctccagt agttgctcct gttcaagtat ctactcctgt tgattctggt 5580 ttgttggcaa aagcggaaca agttgtattg gaagtgttag catgcaagac tggttatgaa 5640 actgagttga ttgaattgga tatggaattg gaaactgaac ttggtattga ttctatcaag 5700 agagtagaaa ttctttctga agttcaagct caattgagtg ttgaagctaa agatgtagat 5760 gctcttagta gaactcgtac tgttggtgaa gtgattgatg caatgaaagc cgaaatttct 5820 ggtggtcaac caactgctcc tgttcaagtt gcagctccta ctc aagtagt tgctcctgtt 5880 aaagtatcta ctcctgttga ttctggtttg ttagcaaagg cggaacaagt agtattggaa 5940 gtattggcat ctaagactgg ttatgaaact gagttgattg aattagatat ggaattggaa 6000 actgaacttg gtattgattc tatcaagaga gtagaaattc tttctgaagt tcaagctcaa 6060 ttgaatgtgg aagctaaaga tgtggatgct cttagtagaa ctcgtactgt tggtgaagtg 6120 attgatgcaa tgaaagccga aattgctggt gatcaacctg ctccagctgt agttccagtt 6180

130130

201038734 caagctaaga gtggtgtagc caaccctgca cttttggcaa aggcggaaca agtagtattg gaagtattgg catccaagac cggttatgaa actgagctga ttgaattgga tatggaattg gaaactgaac ttggtattga ttcaatcaag agagtagaaa ttctgtccga agttcaagca gaattgagtg ttgaagcaaa agatgtagac gctctaagta gaacccgtac tgttggggaa gtgatcgatg caatgaaagc tgaaattgct ggcagtgctg tcacggttgc aactttggat gattcaacaa ttatggagga gacagatgat gaagatgaag actttatttt atacgatcat gtatacggaa gcgaatgtga agatcttagt ctgagttttt catccgtaaa gagcatcccg cgcgctgata aacttttgtt ggataacatt gctgaaaggc caattgttat tgtggattgt ggaacaaagc ttacaactga acttgcaaaa gctattggag aacgtgccgt ggttgctaca ttcagtgcac agagcttggt atcccgtgga ttcgttggta aatcatttac tctaggaaat acagaagaaa gtgagatcga aaagatggtt tcaagcattg aatcttcgta tggaaaaatt ggtggctttg tttatcaaca ttttcatgat agcgactatg gtatgcaact tggatgggcg ttaatggcag cgaaacattt gaaagagtcc ctcaacgacc cgattaagaa tggaagaacc ttctttttgg ctgttgcgcg tatgaatggt aaacttggta tggacaatgc ttcagttcat gatcaaggaa tagtggaatc atgcggtatc gccgaacgtg gtgctatctt tggtttgtgc aaaactttgg atttggaatg gcctaatgtt tttgctcgtg gtgttgatat tgctgaaggt atgagttata gtttggctgc ggaattgatt gttgatgaga tttcttgtgc aaatctttcc attcgggaat ctggttacac gattagcgga gaaagattca caactgaagc tcacaaattg gttactggaa agcctcatgc tccgattaag aagaaggatg ctttcctagt atctggtggt gctcgtggta ttactccact ttgtattcgt gaaattgcta aagcagtgaa aggtggcact tacattttga tgggtcgatc agctttggct gatgaaccct tgtgggctaa tggtaaatcc ggaaaagatt tagataaagc tggtttggca tttttgaagg aagagtttgc agctgggcgt ggtagtaaac caactccaaa agttcacaaa tctttgattg ataaagtgct cggtattagg gaggttagag catctattgc aaatatagaa gcccatggag caaaagctat atatttgtct tgcgatgtat cttccgctga gaaagtaaag gctgcagtgc aaaaagttga aaaggagcat ctagttcgta ttactggtat tgtgcatgca tcaggcgttt tgagggataa attggttgag aacaaaactt tggatgattt caacgcagta tatggaacca aagtaactgg actagtaaac ttgctgtcag cagtgaacat gaattttgtt cgtcatttgg ttatgtttag ttctttggct ggatatcatg gaaatgttgg tcaatctgat tatgcaatgg ctaacgaatc acttaacaag attggtttta gattgggtgc agcttattct caattgtgtg ttaaatctat ttgttttgga ccttgggatg gtggaatggt aactccagct ttgaaaaaac aatttcaatc aatgggtgtc cagattattc ctcgtgaagg tggcgcggag actgttgcaa gaatagtctt atcttcaaat ccttctcaag ttttagttgg caactggggt gttcctccag tttcaccttt gtcaaaatcg gcaactattg ttcaaacttt tacccctgag ttaaatccat ttctaaagtc tcatcaaatt catggtaaaa atgttttgcc tatgactgta gcaattggat atcttgctca cttggttaag aatttttatg ctggtcatca tttgtgggga gttgaagatg ctcaattgtt cagtggtgtt gtaattgacc atgcggtgca agctcaagtg aaattaacgg aacagagttt ggatgatgat ggcaaggtaa aagttcaagc tgttctgact gcttcaaacg ataatggaaa aatggtacct 6240 6300 6360 6420 6480 6540 6600 6660 6720 6780 6840 6900 6960 7020 7080 7140 7200 7260 7320 7380 7440 7500 7560 7620 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 131 201038734 gcatacaaag cagtgattgt tttgggaaaa acaagtagac ctgcgtttat tttgaaagat 8520 ttttcattgc aagaatctaa ttctcgcagt gctgatgagt tgtatgatgg taaaactttg 8580 tttcatggtc cattatttcg tggaattacc aagttgttga atgtatctga tacttcacta 8640 acaactcaat gtaccaatat tgatttgact gctactgaac gtggtcaatt tgcggatatc 8700 gaacctgtga atccttttat ggcggatgct gcatttcasg ctatgcttgt atgggttaga 8760 aatttaagga atagtgcatc tttaccaaac aattgtgaaa gagtagatat ctataaacca 8820 atagcacctg gtgaaaagta ttacactact ttgcaagctt tgggtaatac ctccggttct 8880 gttctcaagt ctgtatttta tatgcacgat gaacaaggag aagtatttct atctggaaga 8940 gctagtgttg ttgtgaatga caagatggag ttttag 8976 <210> 69 <211> 2991 <212> PRT <213〉 破囊壺菌物種201038734 caagctaaga gtggtgtagc caaccctgca cttttggcaa aggcggaaca agtagtattg gaagtattgg catccaagac cggttatgaa actgagctga ttgaattgga tatggaattg gaaactgaac ttggtattga ttcaatcaag agagtagaaa ttctgtccga agttcaagca gaattgagtg ttgaagcaaa agatgtagac gctctaagta gaacccgtac tgttggggaa gtgatcgatg caatgaaagc tgaaattgct ggcagtgctg tcacggttgc aactttggat gattcaacaa ttatggagga gacagatgat gaagatgaag actttatttt atacgatcat gtatacggaa gcgaatgtga agatcttagt ctgagttttt catccgtaaa gagcatcccg cgcgctgata aacttttgtt ggataacatt gctgaaaggc caattgttat tgtggattgt ggaacaaagc ttacaactga acttgcaaaa gctattggag aacgtgccgt ggttgctaca ttcagtgcac agagcttggt atcccgtgga ttcgttggta aatcatttac tctaggaaat acagaagaaa gtgagatcga aaagatggtt tcaagcattg aatcttcgta tggaaaaatt ggtggctttg tttatcaaca ttttcatgat agcgactatg gtatgcaact tggatgggcg ttaatggcag cgaaacattt gaaagagtcc ctcaacgacc cgattaagaa tggaagaacc ttctttttgg ctgttgcgcg tatgaatggt aaacttggta tggacaatgc ttcagttcat gatcaaggaa tagtggaatc atgcggtatc gccgaacgtg gtgctatctt tggtttgtgc aaaactttgg atttggaatg gcctaatgtt tttgctcgtg gtgttgatat tgctgaaggt atgagttata gtttggctgc ggaattgatt gttgatgaga tttcttgtgc aaatctttcc attcgggaat ctggttacac gattagcgga gaaagattca caactgaagc tcacaaattg gttactggaa agcctcatgc tccgattaag aagaaggatg ctttcctagt atctggtggt gctcgtggta ttactccact ttgtattcgt gaaattgcta aagcagtgaa aggtggcact tacattttga tgggtcgatc agctttggct gatgaaccct tgtgggctaa tggtaaatcc ggaaaagatt tagataaagc tggtttggca tttttgaagg aagagtttgc agctgggcgt ggtagtaaac caactccaaa agttcacaaa tctttgattg ataaagtgct cggtattagg gaggttagag catctattgc aaatatagaa gcccatggag caaaagctat atatttgtct tgcgatgtat cttccgctga gaaagtaaag gctgcagtgc aaaaagttga aaaggagcat ctagttcgta ttactggtat tgtgcatgca tcaggcgttt tgagggataa attggttgag aacaaaactt tggatgattt caacgcagta tatggaacca aagtaactgg actagtaaac ttgctgtcag cagtgaacat gaattttgtt cgtcatttgg ttatgtttag ttctttggct ggatatcatg gaaatgttgg tcaatctgat tatgcaatgg ctaacgaatc acttaacaag attggtttta gattgggtgc agcttattct caattgtgtg ttaaatctat ttgttttgga ccttgggatg gtggaatggt aactccagct ttgaaaaaac aatttcaatc aatgggtgtc cagattattc ctcgtgaagg tggcgcggag actgttgcaa gaatagtctt atcttcaaat ccttctcaag ttttagttgg caactggggt gttcctccag tttcaccttt gtcaaaatcg gcaactattg ttcaaacttt tacccctgag ttaaatccat ttctaaagtc tcatcaaatt catggtaaaa atgttttgcc tatgactgta gcaattggat atcttgctca cttggttaag aatttttatg ctggtcatca tttgtgggga gttgaagatg ctcaattgtt cagtggtgtt gtaattgacc atgcggtgca agctcaagtg aaattaacgg aacagagttt ggatgatgat ggcaaggtaa aagttcaagc tgttctgact gcttcaaacg ataatggaaa aatggtacct 6240 6300 6360 6420 6480 6540 6600 6660 6720 6780 6840 6900 6960 7020 7080 7140 7200 7260 7320 7380 7440 7500 7560 7620 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 131 201038734 gcatacaaag cagtgattgt tttgggaaaa acaagtagac ctgcgtttat tttgaaagat 8520 ttttcattgc aagaatctaa ttctcgcagt gctgatgagt tgtatgatgg taaaactttg 8580 tttcatggtc cattatttcg tggaattacc aagttgttga atgtatctga tacttcacta 8640 acaactcaat gtaccaatat tgatttgact gctactgaac gtggtcaatt tgcggat atc 8700 gaacctgtga atccttttat ggcggatgct gcatttcasg ctatgcttgt atgggttaga 8760 aatttaagga atagtgcatc tttaccaaac aattgtgaaa gagtagatat ctataaacca 8820 atagcacctg gtgaaaagta ttacactact ttgcaagctt tgggtaatac ctccggttct 8880 gttctcaagt ctgtatttta tatgcacgat gaacaaggag aagtatttct atctggaaga 8940 gctagtgttg ttgtgaatga caagatggag ttttag 8976 < 210 > 69 < 211 > 2991 < 212 > PRT <213> Thraustochytrium species

<400> 69<400> 69

Met Glu Asp Gin Arg lie Ala lie Val Gly Leu Ser Ala lie Leu Pro 15 10 15Met Glu Asp Gin Arg lie Ala lie Val Gly Leu Ser Ala lie Leu Pro 15 10 15

Ser Gly Glu Asn Val Arg Glu Ser Trp Glu Ala lie Arg Asp Gly Leu 20 25 30Ser Gly Glu Asn Val Arg Glu Ser Trp Glu Ala lie Arg Asp Gly Leu 20 25 30

Asn Cys Leu Ser Asp Leu Pro Ala Asp Arg Val Asp Val Thr Ala Tyr 35 40 45Asn Cys Leu Ser Asp Leu Pro Ala Asp Arg Val Asp Val Thr Ala Tyr 35 40 45

Tyr Asn Pro Thr Lys Gly Val Lys Asp Lys lie Tyr Cys Lys Arg Gly 50 55 60Tyr Asn Pro Thr Lys Gly Val Lys Asp Lys lie Tyr Cys Lys Arg Gly 50 55 60

Gly Phe lie Pro Glu Tyr Glu Phe Asp Ser Arg Glu Phe Gly Leu Asn 65 70 75 80Gly Phe lie Pro Glu Tyr Glu Phe Asp Ser Arg Glu Phe Gly Leu Asn 65 70 75 80

Met Leu Gin Met Glu Asp Ser Asp Ala Asn Gin Thr Leu Thr Leu Leu 85 90 95Met Leu Gin Met Glu Asp Ser Asp Ala Asn Gin Thr Leu Thr Leu Leu 85 90 95

Lys Val Lys Glu Ala Leu Asp Asp Ala Asn lie Pro Ala Phe Thr Asn 100 105 110Lys Val Lys Glu Ala Leu Asp Asp Ala Asn lie Pro Ala Phe Thr Asn 100 105 110

Glu Lys Lys Asn lie Gly Cys Val Leu Gly lie Gly Gly Gly Gin Lys 115 120 125Glu Lys Lys Asn lie Gly Cys Val Leu Gly lie Gly Gly Gly Gin Lys 115 120 125

Ala Ser His Glu Phe Tyr Ser Arg Leu Asn Tyr Val Val Val Asp Lys 130 135 140Ala Ser His Glu Phe Tyr Ser Arg Leu Asn Tyr Val Val Val Asp Lys 130 135 140

Val Leu Arg Lys Met Gly Leu Pro Asp Glu Asp Val Glu Thr Ala Val 145 150 155 160Val Leu Arg Lys Met Gly Leu Pro Asp Glu Asp Val Glu Thr Ala Val 145 150 155 160

Glu Lys Phe Lys Ala Asn Phe Pro Glu Trp Arg Leu Asp Ser Phe Pro 165 170 175Glu Lys Phe Lys Ala Asn Phe Pro Glu Trp Arg Leu Asp Ser Phe Pro 165 170 175

Gly Phe Leu Gly Asn Val Thr Ala Gly Arg Cys Thr Asn Thr Phe Asn 180 185 190Gly Phe Leu Gly Asn Val Thr Ala Gly Arg Cys Thr Asn Thr Phe Asn 180 185 190

Met Glu Gly Met Asn Cys Val Val Asp Ala Ala Cys Ala Ser Ser Leu 132 201038734 195 200 205 lie Ala lie Lys Val Ala 工le Asp Glu Leu Leu His Gly Asp Cvs Asp 210 215 220 r o y4 T 2Met Glu Gly Met Asn Cys Val Val Asp Ala Ala Cys Ala Ser Ser Leu 132 201038734 195 200 205 lie Ala lie Lys Val Ala work le Asp Glu Leu Leu His Gly Asp Cvs Asp 210 215 220 r o y4 T 2

Ala Met lie Ala Gly Ma Thr Cys Thr Asp Asn Ala Leu Gly Met 225 230 235Ala Met lie Ala Gly Ma Thr Cys Thr Asp Asn Ala Leu Gly Met 225 230 235

Met Ala Phe Ser Lys Thr Pro Val Phe Ser Thr Asp Gin Ser Cys Leu 245 250 255Met Ala Phe Ser Lys Thr Pro Val Phe Ser Thr Asp Gin Ser Cys Leu 245 250 255

Ala Tyr Asp Glu Lys Thr Lys Gly Met Leu lie Gly Glu Gly Ser Ala 260 265 270Ala Tyr Asp Glu Lys Thr Lys Gly Met Leu lie Gly Glu Gly Ser Ala 260 265 270

Met Phe Val Leu Lys Arg Tyr Ala Asp Ala Val Arg Asp Gly Asp Thr 275 280 285 oMet Phe Val Leu Lys Arg Tyr Ala Asp Ala Val Arg Asp Gly Asp Thr 275 280 285 o

Val His Ala Val lie Arg Ser Cys Ser Ser Ser Ser Asp Gly Lvs Ala 290 295 300Val His Ala Val lie Arg Ser Cys Ser Ser Ser Ser Asp Gly Lvs Ala 290 295 300

Ser Gly lie Tyr Thr Pro Thr lie Ser Gly Gin Glu Glu Ala lie Leu 305 310 315 320Ser Gly lie Tyr Thr Pro Thr lie Ser Gly Gin Glu Glu Ala lie Leu 305 310 315 320

Arg Ala Tyr Arg Arg Ala Gly Val Ser Pro Asn Thr He Thr Leu Val 325 330 335Arg Ala Tyr Arg Arg Ala Gly Val Ser Pro Asn Thr He Thr Leu Val 325 330 335

Glu Gly His Gly Thr Gly Thr Pro Val Gly Asp Lys lie Glu Leu Thr 340 345 350Glu Gly His Gly Thr Gly Thr Pro Val Gly Asp Lys lie Glu Leu Thr 340 345 350

Ala Leu Arg Asn Val Phe Asp Lys Ala Tyr Gly Pro Gly His Lys Glu 355 360 365Ala Leu Arg Asn Val Phe Asp Lys Ala Tyr Gly Pro Gly His Lys Glu 355 360 365

G _—_ c a rG _—_ c a r

a V a _—I Aa V a _—I A

s y· L e 5 17 I 3 r e s y _—I Gs y· L e 5 17 I 3 r e s y _—I G

Ser Gin lie Gly His Leu Lys Ala 380Ser Gin lie Gly His Leu Lys Ala 380

Val Ala Gly Cys Ala Gly Leu Val Lys Leu Val Met Ala Leu Lys His 385 390 395 400 〇Val Ala Gly Cys Ala Gly Leu Val Lys Leu Val Met Ala Leu Lys His 385 390 395 400 〇

Lys Thr Leu Pro Gin Ser lie Asn Val Glu Asn Pro Pro Asn Leu Val 405 410 415Lys Thr Leu Pro Gin Ser lie Asn Val Glu Asn Pro Pro Asn Leu Val 405 410 415

Asp Gly Thr Val lie Ser Asp Thr Thr Leu Tyr He Asn Thr Met Asn 420 425 430Asp Gly Thr Val lie Ser Asp Thr Thr Leu Tyr He Asn Thr Met Asn 420 425 430

Arg Pro Trp He Thr Lys Pro Gly Val Pro Arg Arg Ala Gly He Ser 435 440 445Arg Pro Trp He Thr Lys Pro Gly Val Pro Arg Arg Ala Gly He Ser 435 440 445

Ser Phe Gly Phe Gly Gly Ala Asn Tyr His Ala Val Leu Glu Glu Phe 450 455 460Ser Phe Gly Phe Gly Gly Ala Asn Tyr His Ala Val Leu Glu Glu Phe 450 455 460

Glu Pro Glu Gin Thr Lys Pro Tyr Arg Leu Asn Val Ser Ala Gin Pro 465 470 475 480Glu Pro Glu Gin Thr Lys Pro Tyr Arg Leu Asn Val Ser Ala Gin Pro 465 470 475 480

Met Leu Leu His Ala Val Asn Ala Asn Ser Leu Gin Lys Leu Cys Glu 485 490 495 133 201038734Met Leu Leu His Ala Val Asn Ala Asn Ser Leu Gin Lys Leu Cys Glu 485 490 495 133 201038734

Asp Gin Leu Lys Leu Leu Lys Glu Ser Arg Glu Lys Cys Val Asn Thr 500 505 510Asp Gin Leu Lys Leu Leu Lys Glu Ser Arg Glu Lys Cys Val Asn Thr 500 505 510

Lys Asn Thr Asp Tyr Val Ala Phe Ser Lys Phe Gin Asp Ser Phe Lvs 515 520 525Lys Asn Thr Asp Tyr Val Ala Phe Ser Lys Phe Gin Asp Ser Phe Lvs 515 520 525

Leu Lys Gly Ser Val Pro Ser Gin His Ala Arg Val Gly Phe Ala Ser 530 535 540Leu Lys Gly Ser Val Pro Ser Gin His Ala Arg Val Gly Phe Ala Ser 530 535 540

Lys Ser lie Glu Asp Thr lie Ser lie Leu Ser Ala lie Val Asn Arq 545 550 555 560Lys Ser lie Glu Asp Thr lie Ser lie Leu Ser Ala lie Val Asn Arq 545 550 555 560

Phe Gin Lys Asp lie Thr Thr Thr Ser Trp Ala Leu Pro Lys Glu Gly 565 570 575Phe Gin Lys Asp lie Thr Thr Thr Ser Trp Ala Leu Pro Lys Glu Gly 565 570 575

Ala lie Phe Arg Ser Thr Ala Leu lie Asn Asp Asn Lys Ser Val Ala 580 585 590Ala lie Phe Arg Ser Thr Ala Leu lie Asn Asp Asn Lys Ser Val Ala 580 585 590

Ala Leu Phe Ser Gly Gin Gly Ala Gin Tyr Thr His Met Phe Asn Asp 595 600 605Ala Leu Phe Ser Gly Gin Gly Ala Gin Tyr Thr His Met Phe Asn Asp 595 600 605

Val Ala Met Gin Trp Pro Gin Phe Arg Leu Cys Val Asn Asp Met Glu 610 615 620Val Ala Met Gin Trp Pro Gin Phe Arg Leu Cys Val Asn Asp Met Glu 610 615 620

Lys Ala Gin Glu Glu Val lie Asn Asp Lys Ser Val Lys Arg lie Ser 625 630 635 640Lys Ala Gin Glu Glu Val lie Asn Asp Lys Ser Val Lys Arg lie Ser 625 630 635 640

Gin Val Met Phe Pro Arg Lys Pro Tyr Ala Arg Glu Ser Pro Leu Asp 645 650 655Gin Val Met Phe Pro Arg Lys Pro Tyr Ala Arg Glu Ser Pro Leu Asp 645 650 655

Asn Lys Glu lie Ser Lys Thr Glu Tyr Ser Gin Thr Thr Thr Val Ala 660 665 670Asn Lys Glu lie Ser Lys Thr Glu Tyr Ser Gin Thr Thr Thr Val Ala 660 665 670

Ser Ser Val Gly Leu Phe Glu lie Phe Arg Asp Ala Gly Phe Ala Pro 675 680 685Ser Ser Val Gly Leu Phe Glu lie Phe Arg Asp Ala Gly Phe Ala Pro 675 680 685

Ala Phe Val Ala Gly His Ser Leu Gly Glu Phe Ser Ala Leu Tyr Ala 690 695 700Ala Phe Val Ala Gly His Ser Leu Gly Glu Phe Ser Ala Leu Tyr Ala 690 695 700

Ala Gly Leu lie Asp Arg Glu Asp Leu Phe Lys Leu Val Cys Asn Arg 705 710 715 720Ala Gly Leu lie Asp Arg Glu Asp Leu Phe Lys Leu Val Cys Asn Arg 705 710 715 720

Ala Met Ala Met Arg Asp Ala Pro Lys Lys Ser Ala Asp Gly Ala Met 725 730 735Ala Met Ala Met Arg Asp Ala Pro Lys Lys Ser Ala Asp Gly Ala Met 725 730 735

Ala Ala Val lie Gly Pro Asn Ala Ser Ser lie Lys Leu Ser Ala Pro 740 745 750Ala Ala Val lie Gly Pro Asn Ala Ser Ser lie Lys Leu Ser Ala Pro 740 745 750

Glu Val Trp Val Ala Asn Asn Asn Ser Pro Ser Gin Thr Val lie Thr 755 760 765Glu Val Trp Val Ala Asn Asn Asn Ser Pro Ser Gin Thr Val lie Thr 755 760 765

Gly Ala Asn Ser Gly Val Gin Ala Glu Thr Ser Lys Leu Lys Thr Gin 770 775 780Gly Ala Asn Ser Gly Val Gin Ala Glu Thr Ser Lys Leu Lys Thr Gin 770 775 780

Gly Phe Arg Val Val His Leu Ala Cys Asp Gly Ala Phe His Ser Pro 785 790 795 800 134 201038734Gly Phe Arg Val Val His Leu Ala Cys Asp Gly Ala Phe His Ser Pro 785 790 795 800 134 201038734

His Met Glu Asn Ala Glu Lys Gin Phe Gin Lys Ala Leu Ser Ala Val 805 810 815His Met Glu Asn Ala Glu Lys Gin Phe Gin Lys Ala Leu Ser Ala Val 805 810 815

Lys Phe Asn Lys Pro Thr Gly Ser Ser Pro Lys lie Phe Ser Asn Val 820 825 830Lys Phe Asn Lys Pro Thr Gly Ser Ser Pro Lys lie Phe Ser Asn Val 820 825 830

Thr Gly Gly Val Phe Thr Asp Pro Lys Thr Ala Leu Ser Arg His Met 835 840 845Thr Gly Gly Val Phe Thr Asp Pro Lys Thr Ala Leu Ser Arg His Met 835 840 845

Thr Ser Ser Val Gin Phe Leu Thr Gin lie Lys Asn Met Tyr Ala Ala 850 855 860Thr Ser Ser Val Gin Phe Leu Thr Gin lie Lys Asn Met Tyr Ala Ala 850 855 860

Gly Ala Arg Val Phe lie Glu Phe Gly Pro Lys Gin Val Leu Ser Lys 865 870 875 880Gly Ala Arg Val Phe lie Glu Phe Gly Pro Lys Gin Val Leu Ser Lys 865 870 875 880

Leu Val Asn Glu lie Phe Pro Gly Asp Thr Ser Val Leu Thr Val Ser 885 890 895 ❹Leu Val Asn Glu lie Phe Pro Gly Asp Thr Ser Val Leu Thr Val Ser 885 890 895 ❹

Val Asn Pro Ala Ser Ala Lys Asp Ser Asp lie Gin Leu Arg Gin Ala 900 905 910Val Asn Pro Ala Ser Ala Lys Asp Ser Asp lie Gin Leu Arg Gin Ala 900 905 910

Ala Val Gin Met Ala Val Ala Gly Val Ala Leu Thr Asp Phe Asp Lys 915 920 925Ala Val Gin Met Ala Val Ala Gly Val Ala Leu Thr Asp Phe Asp Lys 915 920 925

Trp Glu Leu Lys Asp Pro Thr Arg Met Lys Glu Phe Pro Arg Lys Lys 930 935 940Trp Glu Leu Lys Asp Pro Thr Arg Met Lys Glu Phe Pro Arg Lys Lys 930 935 940

Thr Thr Leu Thr Leu Ser Ala Ala Thr Tyr Val Ser Lys Lys Thr Leu 945 950 955 960Thr Thr Leu Thr Leu Ser Ala Ala Thr Tyr Val Ser Lys Lys Thr Leu 945 950 955 960

Gin Glu Arg Glu Arg 工le Met Asn Asp Gly Arg Thr Val Ser Cys Val 965 970 975Gin Glu Arg Glu Arg work le Met Asn Asp Gly Arg Thr Val Ser Cys Val 965 970 975

Gin Arg lie Glu Asn Thr Asn Thr Gly Glu Leu Glu Lys Leu Lys Lys 980 985 990Gin Arg lie Glu Asn Thr Asn Thr Gly Glu Leu Glu Lys Leu Lys Lys 980 985 990

Gin Leu Gin Asp Lys Glu Asn Glu Val Val Arg Val Gin Ala Leu Ala 995 1000 1005Gin Leu Gin Asp Lys Glu Asn Glu Val Val Arg Val Gin Ala Leu Ala 995 1000 1005

Thr Gin Ala Ser Ala Asp Leu Gin Asn Thr Lys Ala Glu Leu Gin 1010 1015 1020Thr Gin Ala Ser Ala Asp Leu Gin Asn Thr Lys Ala Glu Leu Gin 1010 1015 1020

Lys Ala Gin Ala Thr Lys Ser Ser Asn Ala Ala Ser Asp Ala Val 1025 1030 1035Lys Ala Gin Ala Thr Lys Ser Ser Asn Ala Ala Ser Asp Ala Val 1025 1030 1035

Val Ala Lys His Lys Ala lie Leu Leu Ala Met Leu Glu Glu Leu 1040 1045 1050Val Ala Lys His Lys Ala lie Leu Leu Ala Met Leu Glu Glu Leu 1040 1045 1050

Glu Thr Gly Lys Ala Val Asp Tyr Ser Ser Phe Ser Lys Gly Gin 1055 1060 1065Glu Thr Gly Lys Ala Val Asp Tyr Ser Ser Phe Ser Lys Gly Gin 1055 1060 1065

Val Ala Ser Pro Ala Thr Val Arg Val Val Ser Ala Pro Val Gin 1070 1075 1080Val Ala Ser Pro Ala Thr Val Arg Val Val Ser Ala Pro Val Gin 1070 1075 1080

Ala Ala Ala Pro Val Gin Val Ser Ala Ser Val Asp Ser Gly Leu 1085 1090 1095 135 201038734Ala Ala Ala Pro Val Gin Val Ser Ala Ser Val Asp Ser Gly Leu 1085 1090 1095 135 201038734

Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val Leu Ala Ser Lys 1100 1105 1110Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val Leu Ala Ser Lys 1100 1105 1110

Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met Glu Leu Glu 1115 1120 1125Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met Glu Leu Glu 1115 1120 1125

Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser 1130 1135 1140Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser 1130 1135 1140

Glu Val Gin Ala Gin Leu Asn Val Glu Ala Lys Asp Val Asp Ala 1145 1150 1155Glu Val Gin Ala Gin Leu Asn Val Glu Ala Lys Asp Val Asp Ala 1145 1150 1155

Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 1160 1165 1170Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 1160 1165 1170

Ala Glu 工le Ala Gly Gly Gin Pro Ala Ala Pro Val Gin Val Ala 1175 1180 1185Ala Glu work le Ala Gly Gly Gin Pro Ala Ala Pro Val Gin Val Ala 1175 1180 1185

Ala Pro Thr Gin Val Val Ala Pro Val Gin Ala Ser Ala Pro Val 1190 1195 1200Ala Pro Thr Gin Val Val Ala Pro Val Gin Ala Ser Ala Pro Val 1190 1195 1200

Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 1205 1210 1215Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 1205 1210 1215

Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp 1220 1225 1230Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp 1220 1225 1230

Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val 1235 1240 1245Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val 1235 1240 1245

Glu lie Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys 1250 1255 1260Glu lie Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys 1250 1255 1260

Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie 1265 1270 1275Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie 1265 1270 1275

Asp Ala Met Lys Ala Glu lie Ala Gly Gly Gin Pro Ala Ala Pro 1280 1285 1290Asp Ala Met Lys Ala Glu lie Ala Gly Gly Gin Pro Ala Ala Pro 1280 1285 1290

Val Gin Val Ala Ala Pro Thr Gin Val Val Ala Pro Val Gin Ala 1295 1300 1305Val Gin Val Ala Ala Pro Thr Gin Val Val Ala Pro Val Gin Ala 1295 1300 1305

Ser Ala Pro Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val 1310 1315 1320Ser Ala Pro Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val 1310 1315 1320

Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu 1325 1330 1335 lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser 1340 1345 1350 lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu Ser 1355 1360 1365Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu 1325 1330 1335 lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser 1340 1345 1350 lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu Ser 1355 1360 1365

Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val 136 201038734 1370 1375 1380Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val 136 201038734 1370 1375 1380

Gly Glu Val lie Asp Ala Met Lys Ala Glu lie Ser Gly Gly Gin 1385 1390 1395Gly Glu Val lie Asp Ala Met Lys Ala Glu lie Ser Gly Gly Gin 1385 1390 1395

Pro Ala Ala Pro Val Gin Val Ala Ala Pro Thr Gin lie Val Ala 1400 1405 1410Pro Ala Ala Pro Val Gin Val Ala Ala Pro Thr Gin lie Val Ala 1400 1405 1410

Pro Val Gin Val Ser Ala Pro Val Asp Ser Gly Leu Leu Ala Lys 1415 1420 1425Pro Val Gin Val Ser Ala Pro Val Asp Ser Gly Leu Leu Ala Lys 1415 1420 1425

Ala Glu Gin Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr 1430 1435 1440Ala Glu Gin Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr 1430 1435 1440

Glu Thr Glu Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu 1445 1450 1455 ΟGlu Thr Glu Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu 1445 1450 1455 Ο

Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin 1460 1465 1470Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin 1460 1465 1470

Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg 1475 1480 1485Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg 1475 1480 1485

Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys Ala Glu lie 1490 1495 1500Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys Ala Glu lie 1490 1495 1500

Ser Gly Gly Gin Pro Thr Ala Pro Val Gin Val Ala Ala Pro Thr 1505 1510 1515Ser Gly Gly Gin Pro Thr Ala Pro Val Gin Val Ala Ala Pro Thr 1505 1510 1515

Gin lie Val Ala Pro Val Gin Val Ser Ala Pro Val Asp Ser Gly 1520 1525 1530Gin lie Val Ala Pro Val Gin Val Ser Ala Pro Val Asp Ser Gly 1520 1525 1530

Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val Leu Ala Ser 1535 1540 1545Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val Leu Ala Ser 1535 1540 1545

Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met Glu Leu 1550 1555 1560Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met Glu Leu 1550 1555 1560

Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu 1565 1570 1575Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu 1565 1570 1575

Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 1580 1585 1590Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 1580 1585 1590

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met 1595 1600 1605Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met 1595 1600 1605

Lys Ala Glu He Ser Gly Gly Gin Pro Ala Ala Pro Val Gin Val 1610 1615 1620Lys Ala Glu He Ser Gly Gly Gin Pro Ala Ala Pro Val Gin Val 1610 1615 1620

Ala Ala Pro Thr Gin He Val Ala Pro Val Gin Ala Ser Ala Pro 1625 1630 1635Ala Ala Pro Thr Gin He Val Ala Pro Val Gin Ala Ser Ala Pro 1625 1630 1635

Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu 1640 1645 1650 137 201038734Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu 1640 1645 1650 137 201038734

Val Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu 1655 1660 1665Val Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu 1655 1660 1665

Asp Met Glu Leu Glu Thr Glu Leu Gly 工le Asp Ser lie Lys Arg 1670 1675 1680Asp Met Glu Leu Glu Thr Glu Leu Gly work le Asp Ser lie Lys Arg 1670 1675 1680

Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala 1685 1690 1695Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala 1685 1690 1695

Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val 1700 1705 1710 工le Asp Ala Met Lys Ala Glu lie Ser Gly Gly Gin Pro Ala Ala 1715 1720 1725Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val 1700 1705 1710 work le Asp Ala Met Lys Ala Glu lie Ser Gly Gly Gin Pro Ala Ala 1715 1720 1725

Pro Val Gin Val Ala Ala Pro Thr Gin 工le Val Ala Pro Val Gin 1730 1735 1740Pro Val Gin Val Ala Ala Pro Thr Gin Le Val Ala Pro Val Gin 1730 1735 1740

Val Ser Ala Pro Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin 1745 1750 1755Val Ser Ala Pro Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin 1745 1750 1755

Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu 1760 1765 1770Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu 1760 1765 1770

Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp 1775 1780 1785Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp 1775 1780 1785

Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu 1790 1795 1800Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu 1790 1795 1800

Asn Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr 1805 1810 1815Asn Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr 1805 1810 1815

Val Gly Glu Val He Asp Ala Met Lys Ala Glu lie Ala Gly Gly 1820 1825 1830Val Gly Glu Val He Asp Ala Met Lys Ala Glu lie Ala Gly Gly 1820 1825 1830

Gin Pro Ala Ala Pro Val Gin Val Ala Ala Pro Ala Pro Val Val 1835 1840 1845Gin Pro Ala Ala Pro Val Gin Val Ala Ala Pro Ala Pro Val Val 1835 1840 1845

Ala Pro Val Gin Val Ser Thr Pro Val Asp Ser Gly Leu Leu Ala 1850 1855 I860Ala Pro Val Gin Val Ser Thr Pro Val Asp Ser Gly Leu Leu Ala 1850 1855 I860

Lys Ala Glu Gin Val Val Leu Glu Val Leu Ala Cys Lys Thr Gly 1865 1870 1875Lys Ala Glu Gin Val Val Leu Glu Val Leu Ala Cys Lys Thr Gly 1865 1870 1875

Tyr Glu Thr Glu Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu 1880 1885 1890Tyr Glu Thr Glu Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu 1880 1885 1890

Leu Gly 工le Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val 1895 1900 1905Leu Gly work Le Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val 1895 1900 1905

Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp Ala Leu Ser 1910 1915 1920Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp Ala Leu Ser 1910 1915 1920

Arg Thr Arg Thr Val Gly Glu Val 工le Asp Ala Met Lys Ala Glu 1925 1930 1935 138 201038734Arg Thr Arg Thr Val Gly Glu Val work asp Ala Met Lys Ala Glu 1925 1930 1935 138 201038734

lie Ser 1940 Gly Gly Gin Pro Thr 1945 Ala Pro Val Gin Val 1950 Ala Ala Pro Thr Gin 1955 Val Val Ala Pro Val 1960 Lys Val Ser Thr Pro 1965 Val Asp Ser Gly Leu 1970 Leu Ala Lys Ala Glu 1975 Gin Val Val Leu Glu 1980 Val Leu Ala Ser Lys 1985 Thr Gly Tyr Glu Thr 1990 Glu Leu lie Glu Leu 1995 Asp Met Glu Leu Glu 2000 Thr Glu Leu Gly He 2005 Asp Ser lie Lys Arg 2010 Val Glu lie Leu Ser 2015 Glu Val Gin Ala Gin 2020 Leu Asn Val Glu Ala 2025 Lys Asp Val Asp Ala 2030 Leu Ser Arg Thr Arg 2035 Thr Val Gly Glu Val 2040 lie Asp Ala Met Lys 2045 Ala Glu lie Ala Gly 2050 Asp Gin Pro Ala Pro 2055 Ala Val Val Pro Val 2060 Gin Ala Lys Ser Gly 2065 Val Ala Asn Pro Ala 2070 Leu Leu Ala Lys Ala 2075 Glu Gin Val Val Leu 2080 Glu Val Leu Ala Ser 2085 Lys Thr Gly Tyr Glu 2090 Thr Glu Leu lie Glu 2095 Leu Asp Met Glu Leu 2100 Glu Thr Glu Leu Gly 2105 He Asp Ser lie Lys 2110 Arg Val Glu lie Leu 2115 Ser Glu Val Gin Ala 2120 Glu Leu Ser Val Glu 2125 Ala Lys Asp Val Asp 2130 Ala Leu Ser Arg Thr 2135 Arg Thr Val Gly Glu 2140 Val lie Asp Ala Met 2145 Lys Ala Glu lie Ala 2150 Gly Ser Ala Val Thr 2155 Val Ala Thr Leu Asp 2160 Asp Ser Thr lie Met 2165 Glu Glu Thr Asp Asp 2170 Glu Asp Glu Asp Phe 2175 lie Leu Tyr Asp His 2180 Val Tyr Gly Ser Glu 2185 Cys Glu Asp Leu Ser 2190 Leu Ser Phe Ser Ser 2195 Val Lys Ser lie Pro 2200 Arg Ala Asp Lys Leu 2205 Leu Leu Asp Asn lie 2210 Ala Glu Arg Pro lie 2215 Val lie Val Asp Cys 2220 Gly Thr Lys 139 201038734Lie Ser 1940 Gly Gly Gin Pro Thr 1945 Ala Pro Val Gin Val 1950 Ala Ala Pro Thr Gin 1955 Val Val Ala Pro Val 1960 Lys Val Ser Thr Pro 1965 Val Asp Ser Gly Leu 1970 Leu Ala Lys Ala Glu 1975 Gin Val Val Leu Glu 1980 Val Leu Ala Ser Lys 1985 Thr Gly Tyr Glu Thr 1990 Glu Leu lie Glu Leu 1995 Asp Met Glu Leu Glu 2000 Thr Glu Leu Gly He 2005 Asp Ser lie Lys Arg 2010 Val Glu lie Leu Ser 2015 Glu Val Gin Ala Gin 2020 Leu Asn Val Glu Ala 2025 Lys Asp Val Asp Ala 2030 Leu Ser Arg Thr Arg 2035 Thr Val Gly Glu Val 2040 lie Asp Ala Met Lys 2045 Ala Glu lie Ala Gly 2050 Asp Gin Pro Ala Pro 2055 Ala Val Val Pro Val 2060 Gin Ala Lys Ser Gly 2065 Val Ala Asn Pro Ala 2070 Leu Leu Ala Lys Ala 2075 Glu Gin Val Val Leu 2080 Glu Val Leu Ala Ser 2085 Lys Thr Gly Tyr Glu 2090 Thr Glu Leu lie Glu 2095 Leu Asp Met Glu Leu 2100 Glu Thr Glu Leu Gly 2105 He Asp Ser lie Lys 2110 Arg Val Glu lie Leu 2115 Ser Glu Val Gin Ala 2120 Glu Leu Ser Val Glu 2125 Ala Lys Asp Val Asp 2130 Ala Leu Ser Arg Thr 2135 A Rg Thr Val Gly Glu 2140 Val lie Asp Ala Met 2145 Lys Ala Glu lie Ala 2150 Gly Ser Ala Val Thr 2155 Val Ala Thr Leu Asp 2160 Asp Ser Thr lie Met 2165 Glu Glu Thr Asp Asp 2170 Glu Asp Glu Asp Phe 2175 lie Leu Tyr Asp His 2180 Val Tyr Gly Ser Glu 2185 Cys Glu Asp Leu Ser 2190 Leu Ser Phe Ser Ser 2195 Val Lys Ser lie Pro 2200 Arg Ala Asp Lys Leu 2205 Leu Leu Asp Asn lie 2210 Ala Glu Arg Pro lie 2215 Val lie Val Asp Cys 2220 Gly Thr Lys 139 201038734

Leu Thr Thr Glu Leu Ala Lys Ala lie Gly Glu Arg Ala Val Val 2225 2230 2235Leu Thr Thr Glu Leu Ala Lys Ala lie Gly Glu Arg Ala Val Val 2225 2230 2235

Ala Thr Phe Ser Ala Gin Ser Leu Val Ser Arg Gly Phe Val Gly 2240 2245 2250Ala Thr Phe Ser Ala Gin Ser Leu Val Ser Arg Gly Phe Val Gly 2240 2245 2250

Lys Ser Phe Thr Leu Gly Asn Thr Glu Glu Ser Glu lie Glu Lys 2255 2260 2265Lys Ser Phe Thr Leu Gly Asn Thr Glu Glu Ser Glu lie Glu Lys 2255 2260 2265

Met Val Ser Ser lie Glu Ser Ser Tyr Gly Lys lie Gly Gly Phe 2270 2275 2280Met Val Ser Ser lie Glu Ser Ser Tyr Gly Lys lie Gly Gly Phe 2270 2275 2280

Val Tyr Gin His Phe His Asp Ser Asp Tyr Gly Met Gin Leu Gly 2285 2290 2295Val Tyr Gin His Phe His Asp Ser Asp Tyr Gly Met Gin Leu Gly 2285 2290 2295

Trp Ala Leu Met Ala Ala Lys His Leu Lys Glu Ser Leu Asn Asp 2300 2305 2310Trp Ala Leu Met Ala Ala Lys His Leu Lys Glu Ser Leu Asn Asp 2300 2305 2310

Pro lie Lys Asn Gly Arg Thr Phe Phe Leu Ala Val Ala Arg Met 2315 2320 2325Pro lie Lys Asn Gly Arg Thr Phe Phe Leu Ala Val Ala Arg Met 2315 2320 2325

Asn Gly Lys Leu Gly Met Asp Asn Ala Ser Val His Asp Gin Gly 2330 2335 2340 lie Val Glu Ser Cys Gly lie Ala Glu Arg Gly Ala lie Phe Gly 2345 2350 2355Asn Gly Lys Leu Gly Met Asp Asn Ala Ser Val His Asp Gin Gly 2330 2335 2340 lie Val Glu Ser Cys Gly lie Ala Glu Arg Gly Ala lie Phe Gly 2345 2350 2355

Leu Cys Lys Thr Leu Asp Leu Glu Trp Pro Asn Val Phe Ala Arg 2360 2365 2370Leu Cys Lys Thr Leu Asp Leu Glu Trp Pro Asn Val Phe Ala Arg 2360 2365 2370

Gly Val Asp lie Ala Glu Gly Met Ser Tyr Ser Leu Ala Ala Glu 2375 2380 2385Gly Val Asp lie Ala Glu Gly Met Ser Tyr Ser Leu Ala Ala Glu 2375 2380 2385

Leu lie Val Asp Glu lie Ser Cys Ala Asn Leu Ser lie Arg Glu 2390 2395 2400Leu lie Val Asp Glu lie Ser Cys Ala Asn Leu Ser lie Arg Glu 2390 2395 2400

Ser Gly Tyr Thr He Ser Gly Glu Arg Phe Thr Thr Glu Ala His 2405 2410 2415Ser Gly Tyr Thr He Ser Gly Glu Arg Phe Thr Thr Glu Ala His 2405 2410 2415

Lys Leu Val Thr Gly Lys Pro His Ala Pro lie Lys Lys Lys Asp 2420 2425 2430Lys Leu Val Thr Gly Lys Pro His Ala Pro lie Lys Lys Lys Ass 2420 2425 2430

Ala Phe Leu Val Ser Gly Gly Ala Arg Gly lie Thr Pro Leu Cys 2435 2440 2445 lie Arg Glu lie Ala Lys Ala Val Lys Gly Gly Thr Tyr lie Leu 2450 2455 2460Ala Phe Leu Val Ser Gly Gly Ala Arg Gly lie Thr Pro Leu Cys 2435 2440 2445 lie Arg Glu lie Ala Lys Ala Val Lys Gly Gly Thr Tyr lie Leu 2450 2455 2460

Met Gly Arg Ser Ala Leu Ala Asp Glu Pro Leu Trp Ala Asn Gly 2465 2470 2475Met Gly Arg Ser Ala Leu Ala Asp Glu Pro Leu Trp Ala Asn Gly 2465 2470 2475

Lys Ser Gly Lys Asp Leu Asp Lys Ala Gly Leu Ala Phe Leu Lys 2480 2485 2490Lys Ser Gly Lys Asp Leu Asp Lys Ala Gly Leu Ala Phe Leu Lys 2480 2485 2490

Glu Glu Phe Ala Ala Gly Arg Gly Ser Lys Pro Thr Pro Lys Val 140 201038734Glu Glu Phe Ala Ala Gly Arg Gly Ser Lys Pro Thr Pro Lys Val 140 201038734

His 2495 2500 2505 Lys Ser Leu lie Asp Lys Val Leu Gly lie Arg Glu Val Arg 2510 2515 2520 Ala Ser lie Ala Asn lie Glu Ala His Gly Ala Lys Ala lie Tyr 2525 2530 2535 Leu Ser Cys Asp Val Ser Ser Ala Glu Lys Val Lys Ala Ala Val 2540 2545 2550 Gin Lys Val Glu Lys Glu His Leu Val Arg lie Thr Gly lie Val 2555 2560 2565 His Ala Ser Gly Val Leu Arg Asp Lys Leu Val Glu Asn Lys Thr 2570 2575 2580 Leu ❹ Asp Asp Phe Asn Ala Val Tyr Gly Thr Lys Val Thr Gly Leu 2585 2590 2595 Val Asn Leu Leu Ser Ala Val Asn Met Asn Phe Val Arg His Leu 2600 2605 2610 Val Met Phe Ser Ser Leu Ala Gly Tyr His Gly Asn Val Gly Gin 2615 2620 2625 · Ser Asp Tyr Ala Met Ala Asn Glu Ser Leu Asn Lys lie Gly Phe 2630 2635 2640 - Arg Leu Gly Ala Ala Tyr Ser Gin Leu Cys Val Lys Ser lie Cys 2645 2650 2655 Phe Gly Pro Trp Asp Gly Gly Met Val Thr Pro Ala Leu Lys Lys 2660 2665 2670 Gin Phe Gin Ser Met Gly Val Gin lie lie Pro Arg Glu Gly Gly 2675 2680 2685 O Ala Glu Thr Val Ala Arg lie Val Leu Ser Ser Asn Pro Ser Gin 2690 2695 2700 Val Leu Val Gly Asn Trp Gly Val Pro Pro Val Ser Pro Leu Ser 2705 2710 2715 Lys Ser Ala Thr lie Val Gin Thr Phe Thr Pro Glu Leu Asn Pro 2720 2725 2730 Phe Leu Lys Ser His Gin lie His Gly Lys Asn Val Leu Pro Met 2735 2740 2745 Thr Val Ala lie Gly Tyr Leu Ala His Leu Val Lys Asn Phe Tyr 2750 2755 2760His 2495 2500 2505 Lys Ser Leu lie Asp Lys Val Leu Gly lie Arg Glu Val Arg 2510 2515 2520 Ala Ser lie Ala Asn lie Glu Ala His Gly Ala Lys Ala lie Tyr 2525 2530 2535 Leu Ser Cys Asp Val Ser Ser Ala Glu Lys Val Lys Ala Ala Val 2540 2545 2550 Gin Lys Val Glu Lys Glu His Leu Val Arg lie Thr Gly lie Val 2555 2560 2565 His Ala Ser Gly Val Leu Arg Asp Lys Leu Val Glu Asn Lys Thr 2570 2575 2580 Leu ❹ Asp Asp Phe Asn Ala Val Tyr Gly Thr Lys Val Thr Gly Leu 2585 2590 2595 Val Asn Leu Leu Ser Ala Val Asn Met Asn Phe Val Arg His Leu 2600 2605 2610 Val Met Phe Ser Ser Leu Ala Gly Tyr His Gly Asn Val Gly Gin 2615 2620 2625 · Ser Asp Tyr Ala Met Ala Asn Glu Ser Leu Asn Lys lie Gly Phe 2630 2635 2640 - Arg Leu Gly Ala Ala Tyr Ser Gin Leu Cys Val Lys Ser lie Cys 2645 2650 2655 Phe Gly Pro Trp Asp Gly Gly Met Val Thr Pro Ala Leu Lys Lys 2660 2665 2670 Gin Phe Gin Ser Met Gly Val Gin lie lie Pro Arg Glu Gly Gly 2675 2680 2685 O Ala Glu Thr Val Ala Arg lie Val Leu Ser Ser Asn Pro Ser Gin 26 90 2695 2700 Val Leu Val Gly Asn Trp Gly Val Pro Pro Val Ser Pro Leu Ser 2705 2710 2715 Lys Ser Ala Thr lie Val Gin Thr Phe Thr Pro Glu Leu Asn Pro 2720 2725 2730 Phe Leu Lys Ser His Gin lie His Gly Lys Asn Val Leu Pro Met 2735 2740 2745 Thr Val Ala lie Gly Tyr Leu Ala His Leu Val Lys Asn Phe Tyr 2750 2755 2760

Ala Gly His His Leu Trp Gly Val Glu Asp Ala Gin Leu Phe Ser 2765 2770 2775 141 201038734Ala Gly His His Leu Trp Gly Val Glu Asp Ala Gin Leu Phe Ser 2765 2770 2775 141 201038734

Gly Val Val lie Asp His Ala Val Gin Ala Gin Val Lys Leu Thr 2780 2785 2790Gly Val Val lie Asp His Ala Val Gin Ala Gin Val Lys Leu Thr 2780 2785 2790

Glu Gin Ser Leu Asp Asp Asp Gly Lys Val Lys Val Gin Ala Val 2795 2800 2805Glu Gin Ser Leu Asp Asp Asp Gly Lys Val Lys Val Gin Ala Val 2795 2800 2805

Leu Thr Ala Ser Asn Asp Asn Gly Lys Met Val Pro Ala Tyr Lys 2810 2815 2820Leu Thr Ala Ser Asn Asp Asn Gly Lys Met Val Pro Ala Tyr Lys 2810 2815 2820

Ala Val lie Val Leu Gly Lys Thr Ser Arg Pro Ala Phe lie Leu 2825 2830 2835Ala Val lie Val Leu Gly Lys Thr Ser Arg Pro Ala Phe lie Leu 2825 2830 2835

Lys Asp Phe Ser Leu Gin Glu Ser Asn Ser Arg Ser Ala Asp Glu 2840 2845 2850Lys Asp Phe Ser Leu Gin Glu Ser Asn Ser Arg Ser Ala Asp Glu 2840 2845 2850

Leu Tyr Asp Gly Lys Thr Leu Phe His Gly Pro Leu Phe Arg Gly 2855 2860 2865Leu Tyr Asp Gly Lys Thr Leu Phe His Gly Pro Leu Phe Arg Gly 2855 2860 2865

lie Thr Lys Leu Leu Asn Val Ser Asp Thr Ser Leu Thr Thr Gin 2870 2875 2880Lie Thr Lys Leu Leu Asn Val Ser Asp Thr Ser Leu Thr Thr Gin 2870 2875 2880

Cys Thr Asn lie Asp Leu Thr Ala Thr Glu Arg Gly Gin Phe Ala 2885 2890 2895Cys Thr Asn lie Asp Leu Thr Ala Thr Glu Arg Gly Gin Phe Ala 2885 2890 2895

Asp lie Glu Pro Val Asn Pro Phe Met Ala Asp Ala Ala Phe Gin 2900 2905 2910Asp lie Glu Pro Val Asn Pro Phe Met Ala Asp Ala Ala Phe Gin 2900 2905 2910

Ala Met Leu Val Trp Val Arg Asn Leu Arg Asn Ser Ala Ser Leu 2915 2920 2925Ala Met Leu Val Trp Val Arg Asn Leu Arg Asn Ser Ala Ser Leu 2915 2920 2925

Pro Asn Asn Cys Glu Arg Val Asp lie Tyr Lys Pro lie Ala Pro 2930 2935 2940Pro Asn Asn Cys Glu Arg Val Asp lie Tyr Lys Pro lie Ala Pro 2930 2935 2940

Gly Glu Lys Tyr Tyr Thr Thr Leu Gin Ala Leu Gly Asn Thr Ser 2945 2950 2955Gly Glu Lys Tyr Tyr Thr Thr Leu Gin Ala Leu Gly Asn Thr Ser 2945 2950 2955

Gly Ser Val Leu Lys Ser Val Phe Tyr Met His Asp Glu Gin Gly 2960 2965 2970Gly Ser Val Leu Lys Ser Val Phe Tyr Met His Asp Glu Gin Gly 2960 2965 2970

Glu Val Phe Leu Ser Gly Arg Ala Ser Val Val Val Asn Asp Lys 2975 2980 2985Glu Val Phe Leu Ser Gly Arg Ala Ser Val Val Val Asn Asp Lys 2975 2980 2985

Met Glu Phe 2990 <210> 70 <211> 5880 <212> DNA <213> 破囊壺菌物種 <400> 70 atggtgaaat taagtgttgg tgataatatt tgtcatgatc aacgtgttgc tgttgttggt 60 atggctgtta tgtatgctgg ttgtcaaaat caacatgaat tttggcaatc tttacaaggt 120 aaaaatatga attcaaaatc gatttcacaa aatcgtttag gttctgagta tagagaagaa 180 cattttaaac ctgaaagaag taaatattcc gatacctttt gtaatgaaag atatggttgt 240 attgatgaga atgttcaaag tgaacatgaa cttttattaa aacttgcaaa agatgctatt 300 142 201038734Met Glu Phe 2990 <210> 70 <211> 5880 <212> DNA <213> Thraustochytrium species <400> 70 atggtgaaat taagtgttgg tgataatatt tgtcatgatc aacgtgttgc tgttgttggt 60 atggctgtta tgtatgctgg ttgtcaaaat caacatgaat tttggcaatc tttacaaggt 120 aaaaatatga attcaaaatc gatttcacaa Aatcgtttag gttctgagta tagagaagaa 180 cattttaaac ctgaaagaag taaatattcc gatacctttt gtaatgaaag atatggttgt 240 attgatgaga atgttcaaag tgaacatgaa cttttattaa aacttgcaaa agatgctatt 300 142 201038734

gcggatacaa aaggttctat tgatttgaat aaaaccggaa tcgttagtgg ttgcttatct 360 tttccaatgg ataatttaca aggtgattta ttaaatttgt atcaatgtca cattgaaaag 420 aaaattgggc caaatgcatt aaaagatgtg aatttatggt ctaaaagaac caccaacgga 480 aaagatgata aaaaagctta ttttgatcct gcctctttcg tagctgaaca attagatatg 540 ggaccattac attatagttt agatgctgct tgtgcgtctg cactttatgt attaagactt 600 gctcaagatc atttattaag tggtgctgct gatacaatgt tatgtggtgc atcttgttta 660 cctgaacctt tttttatttt atctggtttt tctacttttc atgcaatgcc attatctggt 720 gatgtttctg ctcctttgca taaaacttca caaggtctta cacctggtga aggtggtgct 780 attatggtac ttaaacgatt aaatgatgca atccgtgatg gtgatagaat ttatggtact 840 ttacttggtg ctgaattaag taatgctggt tgtggtttac cattgagtcc acatatgcca 900 agtgaatttg attgtatgga aaaagcttta caaagagtac acagattacc atcatctatt 960 caatatgttg agtgtcatgc aactggtaca ccacaaggtg ataaagttga aattgatgct 1020 atgacaaaat gttttggtga acatttacca aggtttggtt caacgaaagg gaattttggt 1080 catacacttg ttgctgctgg ttttgctggt atgtgtaaag ttttattatc aatgcaatat 1140 ggtgaaatac caccaactcc aggtcttgaa aatccagaca atattatgca tgatttagtt 1200 gttactgaaa caattccatg gcctaataca aatggtgatt tgaaacgtgc atgtttatct 1260 gcttttggat tcggtggtac taatgcacat gctgtatttg aagagtatcg ttcagattta 1320 caagcaaata aaactcttga aaatgaaagt aaaagtcatg aaatcttttc ttcatttaaa 1380 attgctattg ttggtatgga atctgaattt ggtactttga aaggattaca agaatttgaa 1440 cgtgctattt acaatggtgg tcatggtgca tgtgatttac ctgaaaatag atggagattt 1500 cttggagaag ataaagaatt tttacaagct tgtggtttac aaaaattacc aagaggttgt 1560 tatattaaag aagtggaaac tgattttaaa aggttacgtt taccaatgat acaggaggat 1620 attctaagac ctttacagtt gttagctgtt tcgattatcg acagagcact taacgcatct 1680 ggtgttaaac caaatggcaa agttgcagtt ttagttggat taggtactga tcttgaatta 1740 tatcgtcatc gtgctcgtgt tgcattaaag gaacgcctcc aaactgcggt caaagaagat 1800 attcctttac ttgaaaagtt aatgaactat gtcaatgata gaggtacaag tacatcatat 1860 acatcttata ttggaaattt ggttgcaact cgagtttcat cattatgggg ttttactggt 1920 ccatcattca cgattactga aggtgaaaat tccgtatatc gttgtcttga tttgggaaga 1980 tggttcttag ctaatggtga agtagatgct gttgttgttg ccggggttga tttatgtggt 2040 agtgctgaaa atctttttgt aaaatctcgt agaagtaaag tttccacaca aaatgaacca 2100 tttgcaaatt ttgaatcaaa tgctgatgga tattttgctg gagatggttg tggagctttg 2160 gttttgaaac gattgagtga ttgtacggat tcaactgaaa aaatttatgc aacggtggat 2220 tcaattgctg ttggtgatga agttggccca actattaaac aagctttgaa gaatgcatcc 2280 atagcagcga aagatattga actggcagag ctatcagcaa gttcaggcaa acatcattct 2340 ggtagaatca cttgtgaaga tgaactaaat gaactgggtg aaattttcaa tgaaggtata 2400 caaagagttg caattggtag tgtgaaagct aatgttggag atgttggata tgcatctggt 2460 gcagcaagtt taatcaaaac ggctttgtgc ctgtacaacc gatatttacc aaagttacca 2520 143 201038734 aattggaata agccaacgaa agatgttgaa tggtccaaat cattttttgt atgtgaacat 2580 tctagagcat ggttgaaaaa tgttgatgaa aatagacatg ctgtcgtttc tggagtttgc 2640 gaaaatggtt cgtgttatgg aatcgtaatg tctgatgtac aaggacatca tgaagaatcg 2700 aatcttgtta gtttagacaa aaatgaacca aaagtactgg gtatttacgg agattcagtt 27 60 gatgatatcc tagttcagct caacaaatat cttgaaaaat tccttcaaga aactggaacg 2820 gctgcggctg cacaaaaagt taaatcacct acaatagata ttgactccaa tgtgtttgct 2880 gagatgctta atctaccgca ggataaaaac aaaaaatttg cggtcgcatt ggttaccaca 2940 ccaaataaac tccagcgtga aatagaactt gctgtgaagg gtattccacg ttgcgtaaaa 3000 gcaaaaagag attggtgttc tccatctgga agtatttttg cttgtaatcc actcaaaagt 3060 gataatattg catttatgta tggtgaaggc cgaagcccat atgctggact gggatatgat 3120 ttgcatcgaa tttggcctat gctacacgag ttggttaaca atagaactac agaactttgg 3180 gatcaaggtg atagttggta tttacctcga tctagctctg ttgctgaaaa agaaaaagtc 3240 ttcggagatt ttgataagaa tcaaattgaa atgtttagat tgggtatttt tgtatcaatg 3300 tgtttcactg atatggccac tgaacttttg ggtttaaaac ccaaagccgc gtttggttta 3360 agtttgggtg aaatatctat gctttttgca ttttctaaaa agaataccaa gttgtccaaa 3420 gaattgaccc gtcgtctaaa agaagcaaaa gtttgggcat cacaattagc tgttgaattt 3480 gcagctattc gagatttgtg gaatattcca gctgataaat ctattgatga attttggcaa 3540 gggtattttg tttacgcaaa tcgaaccctg gtcgagaaca caattgggga gaataaattt 3600 gttcgtttgt tgattgtaaa tgattcgcaa agttgtctaa ttgccgggaa accagatgaa 3660 tgtcaaaaag ttattgagaa gcttcatttg aagctaccgg cggttccagt aactcagggt 3720 atgatcggtc attgcccaga agcaattcct tatctagatc aaatcagtca tattcatgaa 3780 atgcttgaaa ttccaaaacc cgaaaatgtg aaattgttta caactagtga aaacagagaa 3840 ttagtgtcga tgaaagattc cgtgtcaaaa ttggttgctg agatttatca gcatgttgct 3900 gattttccaa acatcgtgaa caaggttaaa gaaacttgca aaactgatat atttattgaa 3960 ttgggatcga acaattatcg atctggagct gtcaaaacaa ttttaggtcc agaaatcgtt 4020 tctgttgcaa ttgataggca aaatgaaact gcatggggtc aactaatgaa gatggttgca 4080 tcgttgataa gtcatcgagt tccgggtgtt gaattgaaaa aactctatca tcctgaattg 4140 ctgaaatttg atccacaggc aaaaccgaat cgtttcatca gaaatataga actgaatgga 4200 ttttttgatc gtacgaatat tattgttgat aagcaactat cccctgcgga tccgaaactc 4260 gctgaaattg tgaacaatcg aaatatgcct aaagataatg tttatgtacc aattgaacgg 4320 gtgaaaacga tgataaaggc ggaaccagct aatttacaag tcagcgtggg aagtaaacca 4380 gttgttactg aaagaattag ttcggacgat aatctatttg aaaagttgtc agaaattaca 4440 aaatcttttg atggtgtaaa tgcgtgtact gaagcaatgt tgggagactc tggatttctc 4500 aaaacatatg aggttgacta tcctttgtac acaggtgcca tggctaaagg aattgcgtct 4560 gctgatttgg ttattgctgc tggtaaatca aagatcttgg catcatttgg agctggtggg 4620 ttggccttac aagtggtaga agatgccatt aaacaaatta aagctgaatt ggggaacggt 4680 ccgtttgctg taaatttgat tcattcacca ttcgatccta gcttggagaa gggtaacgtt 4740 gatctttttc taaaatataa cgttcgattt gttgaagtat ccgcatttat gtcattaacc 4800gcggatacaa aaggttctat tgatttgaat aaaaccggaa tcgttagtgg ttgcttatct 360 tttccaatgg ataatttaca aggtgattta ttaaatttgt atcaatgtca cattgaaaag 420 aaaattgggc caaatgcatt aaaagatgtg aatttatggt ctaaaagaac caccaacgga 480 aaagatgata aaaaagctta ttttgatcct gcctctttcg tagctgaaca attagatatg 540 ggaccattac attatagttt agatgctgct tgtgcgtctg cactttatgt attaagactt 600 gctcaagatc atttattaag tggtgctgct gatacaatgt tatgtggtgc atcttgttta 660 cctgaacctt tttttatttt atctggtttt tctacttttc atgcaatgcc attatctggt 720 gatgtttctg ctcctttgca taaaacttca caaggtctta cacctggtga aggtggtgct 780 attatggtac ttaaacgatt aaatgatgca atccgtgatg gtgatagaat ttatggtact 840 ttacttggtg ctgaattaag taatgctggt tgtggtttac cattgagtcc acatatgcca 900 agtgaatttg attgtatgga aaaagcttta caaagagtac acagattacc atcatctatt 960 caatatgttg agtgtcatgc aactggtaca ccacaaggtg ataaagttga aattgatgct 1020 atgacaaaat gttttggtga acatttacca aggtttggtt caacgaaagg gaattttggt 1080 catacacttg ttgctgctgg ttttgctggt atgtgtaaag ttttattatc aatgcaatat 1140 ggt gaaatac caccaactcc aggtcttgaa aatccagaca atattatgca tgatttagtt 1200 gttactgaaa caattccatg gcctaataca aatggtgatt tgaaacgtgc atgtttatct 1260 gcttttggat tcggtggtac taatgcacat gctgtatttg aagagtatcg ttcagattta 1320 caagcaaata aaactcttga aaatgaaagt aaaagtcatg aaatcttttc ttcatttaaa 1380 attgctattg ttggtatgga atctgaattt ggtactttga aaggattaca agaatttgaa 1440 cgtgctattt acaatggtgg tcatggtgca tgtgatttac ctgaaaatag atggagattt 1500 cttggagaag ataaagaatt tttacaagct tgtggtttac aaaaattacc aagaggttgt 1560 tatattaaag aagtggaaac tgattttaaa aggttacgtt taccaatgat acaggaggat 1620 attctaagac ctttacagtt gttagctgtt tcgattatcg acagagcact taacgcatct 1680 ggtgttaaac caaatggcaa agttgcagtt ttagttggat taggtactga tcttgaatta 1740 tatcgtcatc gtgctcgtgt tgcattaaag gaacgcctcc aaactgcggt caaagaagat 1800 attcctttac ttgaaaagtt aatgaactat gtcaatgata gaggtacaag tacatcatat 1860 acatcttata ttggaaattt ggttgcaact cgagtttcat cattatgggg ttttactggt 1920 ccatcattca cgattactga aggtgaaaat tccgtatatc gttgtcttga tttgggaaga 1 980 tggttcttag ctaatggtga agtagatgct gttgttgttg ccggggttga tttatgtggt 2040 agtgctgaaa atctttttgt aaaatctcgt agaagtaaag tttccacaca aaatgaacca 2100 tttgcaaatt ttgaatcaaa tgctgatgga tattttgctg gagatggttg tggagctttg 2160 gttttgaaac gattgagtga ttgtacggat tcaactgaaa aaatttatgc aacggtggat 2220 tcaattgctg ttggtgatga agttggccca actattaaac aagctttgaa gaatgcatcc 2280 atagcagcga aagatattga actggcagag ctatcagcaa gttcaggcaa acatcattct 2340 ggtagaatca cttgtgaaga tgaactaaat gaactgggtg aaattttcaa tgaaggtata 2400 caaagagttg caattggtag tgtgaaagct aatgttggag atgttggata tgcatctggt 2460 gcagcaagtt taatcaaaac ggctttgtgc ctgtacaacc gatatttacc aaagttacca 2520 143 201038734 aattggaata agccaacgaa agatgttgaa tggtccaaat cattttttgt atgtgaacat 2580 tctagagcat ggttgaaaaa tgttgatgaa aatagacatg ctgtcgtttc tggagtttgc 2640 gaaaatggtt cgtgttatgg aatcgtaatg tctgatgtac aaggacatca tgaagaatcg 2700 aatcttgtta gtttagacaa aaatgaacca aaagtactgg gtatttacgg 27 60 gatgatatcc agattcagtt tagttcagct caacaaatat cttgaaaaat t ccttcaaga aactggaacg 2820 gctgcggctg cacaaaaagt taaatcacct acaatagata ttgactccaa tgtgtttgct 2880 gagatgctta atctaccgca ggataaaaac aaaaaatttg cggtcgcatt ggttaccaca 2940 ccaaataaac tccagcgtga aatagaactt gctgtgaagg gtattccacg ttgcgtaaaa 3000 gcaaaaagag attggtgttc tccatctgga agtatttttg cttgtaatcc actcaaaagt 3060 gataatattg catttatgta tggtgaaggc cgaagcccat atgctggact gggatatgat 3120 ttgcatcgaa tttggcctat gctacacgag ttggttaaca atagaactac agaactttgg 3180 gatcaaggtg atagttggta tttacctcga tctagctctg ttgctgaaaa agaaaaagtc 3240 ttcggagatt ttgataagaa tcaaattgaa atgtttagat tgggtatttt tgtatcaatg 3300 tgtttcactg atatggccac tgaacttttg ggtttaaaac ccaaagccgc gtttggttta 3360 agtttgggtg aaatatctat gctttttgca ttttctaaaa agaataccaa gttgtccaaa 3420 gaattgaccc gtcgtctaaa agaagcaaaa gtttgggcat cacaattagc tgttgaattt 3480 gcagctattc gagatttgtg gaatattcca gctgataaat ctattgatga attttggcaa 3540 gggtattttg tttacgcaaa tcgaaccctg gtcgagaaca caattgggga gaataaattt 3600 gttcgtttgt tgattgtaaa tgattcgcaa agtt gtctaa ttgccgggaa accagatgaa 3660 tgtcaaaaag ttattgagaa gcttcatttg aagctaccgg cggttccagt aactcagggt 3720 atgatcggtc attgcccaga agcaattcct tatctagatc aaatcagtca tattcatgaa 3780 atgcttgaaa ttccaaaacc cgaaaatgtg aaattgttta caactagtga aaacagagaa 3840 ttagtgtcga tgaaagattc cgtgtcaaaa ttggttgctg agatttatca gcatgttgct 3900 gattttccaa acatcgtgaa caaggttaaa gaaacttgca aaactgatat atttattgaa 3960 ttgggatcga acaattatcg atctggagct gtcaaaacaa ttttaggtcc agaaatcgtt 4020 tctgttgcaa ttgataggca aaatgaaact gcatggggtc aactaatgaa gatggttgca 4080 tcgttgataa gtcatcgagt gaattgaaaa aactctatca tcctgaattg 4140 ctgaaatttg atccacaggc aaaaccgaat cgtttcatca gaaatataga actgaatgga 4200 ttttttgatc gtacgaatat tattgttgat aagcaactat cccctgcgga tccgaaactc 4260 gctgaaattg tgaacaatcg aaatatgcct aaagataatg tttatgtacc aattgaacgg 4320 gtgaaaacga tgataaaggc ggaaccagct aatttacaag tcagcgtggg aagtaaacca 4380 gttgttactg aaagaattag ttcggacgat aatctatttg aaaagttgtc agaaattaca 4440 aaatcttttg atggtgtaaa tgcgtgt tccgggtgtt act gaagcaatgt tgggagactc tggatttctc 4500 aaaacatatg aggttgacta tcctttgtac acaggtgcca tggctaaagg aattgcgtct 4560 gctgatttgg ttattgctgc tggtaaatca aagatcttgg catcatttgg agctggtggg 4620 ttggccttac aagtggtaga agatgccatt aaacaaatta aagctgaatt ggggaacggt 4680 ccgtttgctg taaatttgat tcattcacca ttcgatccta gcttggagaa gggtaacgtt 4740 gatctttttc taaaatataa cgttcgattt gttgaagtat ccgcatttat gtcattaacc 4800

i 144 201038734 〇 cctcaggttg attcaaaatc ccagcaccca gcccaacttg gggcatactg agaatttgta ggatgcccga acggtaaatc aaagctacat gaattacaag ttgttcaaaa caaaaagttt aatcgtttac tcgttatgtt ggaagagtgc gcgaaaggat attaataaac ctgaatttta tacgatacag gtattattgc aaaatatttt cattacttgt acaatcgacc aacaataccc aggcagcatt aactttcaaa attcggatgt ttttgaagaa aatacaaatc tcaaaaagtc attctcccga ttcgttggta aggattatca caccatgttt atattttacg actatgagga agccgctggt caaaatttca agatgcattg gccaatggct aattcatgtt aaaacattta tgctgcgttt ggaagcaggt taccatggct aggtactatg gattgaggaa gtttgatgaa aaaaattgaa tttgtcgaag aatttggtgt ggatcctgag tggtgcttgt attagatacg ttggccaaag agaacagagt gttgcggatg gatgatatta ttgttacctt aaagttcgaa gagatgggtg acttgtgact ccagccgcag tttccttcac ttaccagcag gtatgggatg cgtgctgaaa tcttccagat ggtccagcaa attttgggta ttctatcaaa ttaacatact caagagatgg tagcggaact gatctattag ctgtggaagc tgataattca tcggagcagc ctgcatacat atgtacgtaa atatgttcga gtgctaaaaa atgaggtgaa agaccaagaa gagatgcaaa gggctaatac ttgggtcata gttttccaag gactctctca ctgcatcgaa atctgtgaaa gttcttgaaa tcaagaacaa tgattctggt gcaaagaaat tggtggtatt tgcaactgga agtattgaat tcatggtgtt actatacgat aaaacttgag ttactatatt acttaaaatg cggtgaatct taatgatttt tgttgttcag gttgaaatat ttttatttaa 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 〇 <21〇> <211> <212> <213> 71 1959 PRT 破A壺菌物種 <400> 71 Met Val Lys Leu Ser Val Gly Asp Asn lie Cys His Asp Gin Arg Val 15 10 15i 144 201038734 〇cctcaggttg attcaaaatc ccagcaccca gcccaacttg gggcatactg agaatttgta ggatgcccga acggtaaatc aaagctacat gaattacaag ttgttcaaaa caaaaagttt aatcgtttac tcgttatgtt ggaagagtgc gcgaaaggat attaataaac ctgaatttta tacgatacag gtattattgc aaaatatttt cattacttgt acaatcgacc aacaataccc aggcagcatt aactttcaaa attcggatgt ttttgaagaa aatacaaatc tcaaaaagtc attctcccga ttcgttggta aggattatca caccatgttt atattttacg actatgagga agccgctggt caaaatttca agatgcattg gccaatggct aattcatgtt aaaacattta tgctgcgttt ggaagcaggt taccatggct aggtactatg gattgaggaa gtttgatgaa aaaaattgaa tttgtcgaag aatttggtgt ggatcctgag tggtgcttgt attagatacg ttggccaaag agaacagagt gttgcggatg gatgatatta ttgttacctt aaagttcgaa gagatgggtg acttgtgact ccagccgcag tttccttcac ttaccagcag gtatgggatg cgtgctgaaa tcttccagat ggtccagcaa attttgggta ttctatcaaa ttaacatact caagagatgg tagcggaact gatctattag ctgtggaagc tgataattca tcggagcagc ctgcatacat atgtacgtaa atatgttcga gtgctaaaaa atgaggtgaa agaccaagaa gagatgcaaa gggctaatac ttgggtcata gttttccaag gactctctca c tgcatcgaa atctgtgaaa gttcttgaaa tcaagaacaa tgattctggt gcaaagaaat tggtggtatt tgcaactgga agtattgaat tcatggtgtt actatacgat aaaacttgag ttactatatt acttaaaatg cggtgaatct taatgatttt tgttgttcag gttgaaatat ttttatttaa 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 square < 21〇 > < 211 > <212><213> 71 1959 PRT A-cylinder species <400> 71 Met Val Lys Leu Ser Val Gly Asp Asn lie Cys His Asp Gin Arg Val 15 10 15

Ala Val Val Gly Met Ala Val Met Tyr Ala Gly Cys Gin Asn Gin His 20 25 30 Glu Phe Trp Gin Ser Leu Gin Gly Lys Asn Met Asn Ser Lys Ser lie 35 40 45Ala Val Val Gly Met Ala Val Met Tyr Ala Gly Cys Gin Asn Gin His 20 25 30 Glu Phe Trp Gin Ser Leu Gin Gly Lys Asn Met Asn Ser Lys Ser lie 35 40 45

Ser Gin Asn Arg Leu Gly Ser Glu Tyr Arg 50 55Ser Gin Asn Arg Leu Gly Ser Glu Tyr Arg 50 55

0 r p s LY e h p s 1 H u 1 o G 6 u I—I G0 r p s LY e h p s 1 H u 1 o G 6 u I—I G

Glu Arg Ser Lys Tyr Ser Asp Thr Phe Cys Asn Glu Arg Tyr Gly Cys 65 70 75 80 lie Asp Glu Asn Val Gin Ser Glu His Glu Leu Leu Leu Lys Leu Ala 85 90 95Glu Arg Ser Lys Tyr Ser Asp Thr Phe Cys Asn Glu Arg Tyr Gly Cys 65 70 75 80 lie Asp Glu Asn Val Gin Ser Glu His Glu Leu Leu Leu Lys Leu Ala 85 90 95

Lys Asp Ala 工le Ala Asp Thr Lys Gly Ser 工le Asp Leu Asn Lys Thr 100 105 110Lys Asp Ala work le Ala Asp Thr Lys Gly Ser work le Asp Leu Asn Lys Thr 100 105 110

Gly 工le Val Ser Gly Cys Leu Ser Phe Pro Met Asp Asn Leu Gin Gly 115 120 125 145 201038734Gly work le Val Ser Gly Cys Leu Ser Phe Pro Met Asp Asn Leu Gin Gly 115 120 125 145 201038734

Asp Leu Leu Asn Leu Tyr Gin Cys His lie Glu Lys Lys lie Gly Pro 130 135 140Asp Leu Leu Asn Leu Tyr Gin Cys His lie Glu Lys Lys lie Gly Pro 130 135 140

Asn Ala Leu Lys Asp Val Asn Leu Trp Ser Lys Arg Thr Thr Asn Gly 145 150 155 160Asn Ala Leu Lys Asp Val Asn Leu Trp Ser Lys Arg Thr Thr Asn Gly 145 150 155 160

Lys Asp Asp Lys Lys Ala Tyr Phe Asp Pro Ala Ser Phe Val Ala Glu 165 170 175Lys Asp Asp Lys Lys Ala Tyr Phe Asp Pro Ala Ser Phe Val Ala Glu 165 170 175

Gin Leu Asp Met Gly Pro Leu His Tyr Ser Leu Asp Ala Ala Cys Ala 180 185 190Gin Leu Asp Met Gly Pro Leu His Tyr Ser Leu Asp Ala Ala Cys Ala 180 185 190

Ser Ala Leu Tyr Val Leu Arg Leu Ala Gin Asp His Leu Leu Ser Gly 195 200 205Ser Ala Leu Tyr Val Leu Arg Leu Ala Gin Asp His Leu Leu Ser Gly 195 200 205

Ala Ala Asp Thr Met Leu Cys Gly Ala Ser Cys Leu Pro Glu Pro Phe 210 215 220Ala Ala Asp Thr Met Leu Cys Gly Ala Ser Cys Leu Pro Glu Pro Phe 210 215 220

Phe lie Leu Ser Gly Phe Ser Thr Phe His Ala Met Pro Leu Ser Gly 225 230 235 240Phe lie Leu Ser Gly Phe Ser Thr Phe His Ala Met Pro Leu Ser Gly 225 230 235 240

Asp Val Ser Ala Pro Leu His Lys Thr Ser Gin Gly Leu Thr Pro Gly 245 250 255Asp Val Ser Ala Pro Leu His Lys Thr Ser Gin Gly Leu Thr Pro Gly 245 250 255

Glu Gly Gly Ala lie Met Val Leu Lys Arg Leu Asn Asp Ala lie Arg 260 265 270Glu Gly Gly Ala lie Met Val Leu Lys Arg Leu Asn Asp Ala lie Arg 260 265 270

Asp Gly Asp Arg lie Tyr Gly Thr Leu Leu Gly Ala Glu Leu Ser Asn 275 280 285Asp Gly Asp Arg lie Tyr Gly Thr Leu Leu Gly Ala Glu Leu Ser Asn 275 280 285

Ala Gly Cys Gly Leu Pro Leu Ser Pro His Met Pro Ser Glu Phe Asp 290 295 300Ala Gly Cys Gly Leu Pro Leu Ser Pro His Met Pro Ser Glu Phe Asp 290 295 300

Cys Met Glu Lys Ala Leu Gin Arg Val His Arg Leu Pro Ser Ser lie 305 310 315 320Cys Met Glu Lys Ala Leu Gin Arg Val His Arg Leu Pro Ser Ser lie 305 310 315 320

Gin Tyr Val Glu Cys His Ala Thr Gly Thr Pro Gin Gly Asp Lys Val 325 330 335Gin Tyr Val Glu Cys His Ala Thr Gly Thr Pro Gin Gly Asp Lys Val 325 330 335

Glu lie Asp Ala Met Thr Lys Cys Phe Gly Glu His Leu Pro Arg Phe 340 345 350Glu lie Asp Ala Met Thr Lys Cys Phe Gly Glu His Leu Pro Arg Phe 340 345 350

Gly Ser Thr Lys Gly Asn Phe Gly His Thr Leu Val Ala Ala Gly Phe 355 360 365Gly Ser Thr Lys Gly Asn Phe Gly His Thr Leu Val Ala Ala Gly Phe 355 360 365

Ala Gly Met Cys Lys Val Leu Leu Ser Met Gin Tyr Gly Glu lie Pro 370 375 380Ala Gly Met Cys Lys Val Leu Leu Ser Met Gin Tyr Gly Glu lie Pro 370 375 380

Pro Thr Pro Gly Leu Glu Asn Pro Asp Asn lie Met His Asp Leu Val 385 390 395 400Pro Thr Pro Gly Leu Glu Asn Pro Asp Asn lie Met His Asp Leu Val 385 390 395 400

Val Thr Glu Thr lie Pro Trp Pro Asn Thr Asn Gly Asp Leu Lys Arg 405 410 415Val Thr Glu Thr lie Pro Trp Pro Asn Thr Asn Gly Asp Leu Lys Arg 405 410 415

Ala Cys Leu Ser Ala Phe Gly Phe Gly Gly Thr Asn Ala His Ala Val 146 201038734 420 425 430Ala Cys Leu Ser Ala Phe Gly Phe Gly Gly Thr Asn Ala His Ala Val 146 201038734 420 425 430

Phe Glu Glu Tyr Arg Ser Asp Leu Gin 435 440 Ala Asn Lys Thr Leu Glu Asn 445 Glu Ser Lys Ser His Glu lie Phe Ser 450 455 Ser Phe Lys lie Ala lie Val 460 Gly Met Glu Ser Glu Phe Gly Thr Leu 465 470 Lys Gly Leu Gin Glu Phe Glu 475 480Phe Glu Glu Tyr Arg Ser Asp Leu Gin 435 440 Ala Asn Lys Thr Leu Glu Asn 445 Glu Ser Lys Ser His Glu lie Phe Ser 450 455 Ser Phe Lys lie Ala lie Val 460 Gly Met Glu Ser Glu Phe Gly Thr Leu 465 470 Lys Gly Leu Gin Glu Phe Glu 475 480

Arg Ala lie Tyr Asn Gly Gly His Gly Ala Cys Asp Leu Pro Glu Asn 485 490 495Arg Ala lie Tyr Asn Gly Gly His Gly Ala Cys Asp Leu Pro Glu Asn 485 490 495

Arg Trp Arg Phe Leu Gly Glu Asp Lys 500 505 Glu Phe Leu Gin Ala Cys Gly 510 Leu Gin Lys Leu Pro Arg Gly Cys Tyr 〇 515 520 lie Lys Glu Val Glu Thr Asp 525 Phe Lys Arg Leu Arg Leu Pro Met lie 530 535 Gin Glu Asp lie Leu Arg Pro 540 Leu Gin Leu Leu Ala Val Ser lie lie 545 550 Asp Arg Ala Leu Asn Ala Ser 555 560 - Gly Val Lys Pro Asn Gly Lys Val Ala - 565 Val Leu Val Gly Leu Gly Thr 570 575 - Asp Leu Glu Leu Tyr Arg His Arg Ala " 580 585 Arg Val Ala Leu Lys Glu Arg 590 Leu Gin Thr Ala Val Lys Glu Asp lie 595 600 Pro Leu Leu Glu Lys Leu Met 605 Asn Tyr Val Asn Asp Arg Gly Thr Ser 610 615 Thr Ser Tyr Thr Ser Tyr lie 620 Gly Asn Leu Val Ala Thr Arg Val Ser 625 630 Ser Leu Trp Gly Phe Thr Gly 635 640 Pro Ser Phe Thr lie Thr Glu Gly Glu 645 Asn Ser Val Tyr Arg Cys Leu 650 655 Asp Leu Gly Arg Trp Phe Leu Ala Asn 660 665 Gly Glu Val Asp Ala Val Val 670 Val Ala Gly Val Asp Leu Cys Gly Ser 675 680 Ala Glu Asn Leu Phe Val Lys 685 Ser Arg Arg Ser Lys Val Ser Thr Gin 690 695 Asn Glu Pro Phe Ala Asn Phe 700 Glu Ser Asn Ala Asp Gly Tyr Phe Ala 705 710 Gly Asp Gly Cys Gly Ala Leu 715 720 147 201038734Arg Trp Arg Phe Leu Gly Glu Asp Lys 500 505 Glu Phe Leu Gin Ala Cys Gly 510 Leu Gin Lys Leu Pro Arg Gly Cys Tyr 〇515 520 lie Lys Glu Val Glu Thr Asp 525 Phe Lys Arg Leu Arg Leu Pro Met lie 530 535 Gin Glu Asp lie Leu Arg Pro 540 Leu Gin Leu Leu Ala Val Ser lie lie 545 550 Asp Arg Ala Leu Asn Ala Ser 555 560 - Gly Val Lys Pro Asn Gly Lys Val Ala - 565 Val Leu Val Gly Leu Gly Thr 570 575 - Asp Leu Glu Leu Tyr Arg His Arg Ala " 580 585 Arg Val Ala Leu Lys Glu Arg 590 Leu Gin Thr Ala Val Lys Glu Asp lie 595 600 Pro Leu Leu Glu Lys Leu Met 605 Asn Tyr Val Asn Arg Gly Thr Ser 610 615 Thr Ser Tyr Thr Ser Tyr lie 620 Gly Asn Leu Val Ala Thr Arg Val Ser 625 630 Ser Leu Trp Gly Phe Thr Gly 635 640 Pro Ser Phe Thr lie Thr Glu Gly Glu 645 Asn Ser Val Tyr Arg Cys Leu 650 655 Asp Leu Gly Arg Trp Phe Leu Ala Asn 660 665 Gly Glu Val Asp Ala Val Val 670 Val Ala Gly Val Asp Leu Cys Gly Ser 675 680 Ala Glu Asn Leu Phe Val Lys 685 Ser Arg Arg Ser Lys Val Ser Thr Gin 690 695 Asn Glu Pro Phe Ala Asn Phe 700 Glu Ser Asn Ala Asp Gly Tyr Phe Ala 705 710 Gly Asp Gly Cys Gly Ala Leu 715 720 147 201038734

Val Leu Lys Arg Leu Ser Asp Cys Thr Asp Ser Thr Glu Lys lie Tyr 725 730 735Val Leu Lys Arg Leu Ser Asp Cys Thr Asp Ser Thr Glu Lys lie Tyr 725 730 735

Ala Thr Val Asp Ser 工le Ala Val Gly Asp Glu Val Gly Pro Thr lie 740 745 750Ala Thr Val Asp Ser Le Ala Val Gly Asp Glu Val Gly Pro Thr lie 740 745 750

Lys Gin Ala Leu Lys Asn Ala Ser lie Ala Ala Lys Asp lie Glu Leu 755 760 765Lys Gin Ala Leu Lys Asn Ala Ser lie Ala Ala Lys Asp lie Glu Leu 755 760 765

Ala Glu Leu Ser Ala Ser Ser Gly Lys His His Ser Gly Arg lie Thr 770 775 780Ala Glu Leu Ser Ala Ser Ser Gly Lys His His Ser Gly Arg lie Thr 770 775 780

Cys Glu Asp Glu Leu Asn Glu Leu Gly Glu lie Phe Asn Glu Gly lie 785 790 795 800Cys Glu Asp Glu Leu Asn Glu Leu Gly Glu lie Phe Asn Glu Gly lie 785 790 795 800

Gin Arg Val Ala lie Gly Ser Val Lys Ala Asn Val Gly Asp Val Gly 805 810 815Gin Arg Val Ala lie Gly Ser Val Lys Ala Asn Val Gly Asp Val Gly 805 810 815

Tyr Ala Ser Gly Ala Ala Ser Leu lie Lys Thr Ala Leu Cys Leu Tyr 820 825 830Tyr Ala Ser Gly Ala Ala Ser Leu lie Lys Thr Ala Leu Cys Leu Tyr 820 825 830

Asn Arg Tyr Leu Pro Lys Leu Pro Asn Trp Asn Lys Pro Thr Lys Asp 835 840 845Asn Arg Tyr Leu Pro Lys Leu Pro Asn Trp Asn Lys Pro Thr Lys Asp 835 840 845

Val Glu Trp Ser Lys Ser Phe Phe Val Cys Glu His Ser Arg Ala Trp 850 855 860Val Glu Trp Ser Lys Ser Phe Phe Val Cys Glu His Ser Arg Ala Trp 850 855 860

Leu Lys Asn Val Asp Glu Asn Arg His Ala Val Val Ser Gly Val Cys 865 870 875 880Leu Lys Asn Val Asp Glu Asn Arg His Ala Val Val Ser Gly Val Cys 865 870 875 880

Glu Asn Gly Ser Cys Tyr Gly lie Val Met Ser Asp Val Gin Gly His 885 890 895Glu Asn Gly Ser Cys Tyr Gly lie Val Met Ser Asp Val Gin Gly His 885 890 895

His Glu Glu Ser Asn Leu Val Ser Leu Asp Lys Asn Glu Pro Lys Val 900 905 910His Glu Glu Ser Asn Leu Val Ser Leu Asp Lys Asn Glu Pro Lys Val 900 905 910

Leu Gly lie Tyr Gly Asp Ser Val Asp Asp lie Leu Val Gin Leu Asn 915 920 925Leu Gly lie Tyr Gly Asp Ser Val Asp Asp lie Leu Val Gin Leu Asn 915 920 925

Lys Tyr Leu Glu Lys Phe Leu Gin Glu Thr Gly Thr Ala Ala Ala Ala 930 935 940Lys Tyr Leu Glu Lys Phe Leu Gin Glu Thr Gly Thr Ala Ala Ala Ala 930 935 940

Gin Lys Val Lys Ser Pro Thr lie Asp lie Asp Ser Asn Val Phe Ala 945 950 955 960Gin Lys Val Lys Ser Pro Thr lie Asp lie Asp Ser Asn Val Phe Ala 945 950 955 960

Glu Met Leu Asn Leu Pro Gin Asp Lys Asn Lys Lys Phe Ala Val Ala 965 970 975Glu Met Leu Asn Leu Pro Gin Asp Lys Asn Lys Lys Phe Ala Val Ala 965 970 975

Leu Val Thr Thr Pro Asn Lys Leu Gin Arg Glu lie Glu Leu Ala Val 980 985 990Leu Val Thr Thr Pro Asn Lys Leu Gin Arg Glu lie Glu Leu Ala Val 980 985 990

Lys Gly lie Pro Arg Cys Val Lys Ala Lys Arg Asp Trp Cys Ser Pro 995 1000 1005Lys Gly lie Pro Arg Cys Val Lys Ala Lys Arg Asp Trp Cys Ser Pro 995 1000 1005

Ser Gly Ser lie Phe Ala Cys Asn Pro Leu Lys Ser Asp Asn lie 1010 1015 1020 148Ser Gly Ser lie Phe Ala Cys Asn Pro Leu Lys Ser Asp Asn lie 1010 1015 1020 148

Met Tyr Gly Glu 1025 Tyr Asp Leu His Arg lie 1040 Asn Arg Thr Thr Glu Leu 1055 Pro Arg Ser Ser Ser Val 1070 Phe Asp Lys Asn Gin lie 1085 Ser Met Cys Phe Thr Asp 1100 Phe Ala Phe Ser Lys Lys 1130 Arg Arg Leu Lys Glu Ala 1145 Glu Phe Ala Ala lie Arg 1160 Ser lie Asp Glu Phe Trp 1175 Thr Leu Val Glu Asn Thr 1190 Ala Val Pro Val Thr Gin 1235 lie Pro Tyr Leu Asp Gin 1250 lie Pro Lys Pro Glu Asn 1265 Arg Glu Leu Val Ser Met 1280 Glu lie Tyr Gin His Val 1295 Gly 1030 Arg Ser Pro Tyr Ala 1035 Gly Leu Gly Trp 1045 Pro Met Leu His Glu 1050 Leu Val Asn Trp 1060 Asp Gin Gly Asp Ser 1065 Trp Tyr Leu Ala 1075 Glu Lys Glu Lys Val 1080 Phe Gly Asp Glu 1090 Met Phe Arg Leu Gly 1095 lie Phe Val Met 1105 Ala Thr Glu Leu Leu 1110 Gly Leu Lys Leu 1120 Ser Leu Gly Glu lie 1125 Ser Met Leu Asn 1135 Thr Lys Leu Ser Lys 1140 Glu Leu Thr Lys 1150 Val Trp Ala Ser Gin 1155 Leu Ala Val Asp 1165 Leu Trp Asn lie Pro 1170 Ala Asp Lys Gin 1180 Gly Tyr Phe Val Tyr 1185 Ala Asn Arg lie 1195 Gly Glu Asn Lys Phe 1200 Val Arg Leu Gin 1210 Ser Cys Leu lie Ala 1215 Gly Lys Pro lie 1225 Glu Lys Leu His Leu 1230 Lys Leu Pro Gly 1240 Met lie Gly His Cys 1245 Pro Glu Ala lie 1255 Ser His lie His Glu 1260 Met Leu Glu Val 1270 Lys Leu Phe Thr Thr 1275 Ser Glu Asn Lys 1285 Asp Ser Val Ser Lys 1290 Leu Val Ala Ala 1300 Asp Phe Pro Asn lie 1305 Val Asn Lys 201038734Met Tyr Gly Glu 1025 Tyr Asp Leu His Arg lie 1040 Asn Arg Thr Thr Glu Leu 1055 Pro Arg Ser Ser Ser Val 1070 Phe Asp Lys Asn Gin lie 1085 Ser Met Cys Phe Thr Asp 1100 Phe Ala Phe Ser Lys Lys 1130 Arg Arg Leu Lys Glu Ala 1145 Glu Phe Ala Ala lie Arg 1160 Ser lie Asp Glu Phe Trp 1175 Thr Leu Val Glu Asn Thr 1190 Ala Val Pro Val Thr Gin 1235 lie Pro Tyr Leu Asp Gin 1250 lie Pro Lys Pro Glu Asn 1265 Arg Glu Leu Val Ser Met 1280 Glu lie Tyr Gin His Val 1295 Gly 1030 Arg Ser Pro Tyr Ala 1035 Gly Leu Gly Trp 1045 Pro Met Leu His Glu 1050 Leu Val Asn Trp 1060 Asp Gin Gly Asp Ser 1065 Trp Tyr Leu Ala 1075 Glu Lys Glu Lys Val <br><br><br><br><br><br><br><br Val Trp Ala Ser Gin 1155 Leu Ala Val Asp 1165 Leu Trp Asn lie Pro 1170 Ala Asp Lys Gin 1180 Gly Tyr Phe Val Tyr 1185 Ala Asn Arg lie 1195 Gly Glu Asn Lys Phe 1200 Val Arg Leu Gin 1210 Ser Cys Leu lie Ala 1215 Gly Lys Pro lie 1225 Glu Lys Leu His Leu 1230 Lys Leu Pro Gly 1240 Met lie Gly His Cys 1245 Pro Glu Ala lie 1255 Ser His lie His Glu 1260 Met Leu Glu Val 1270 Lys Leu Phe Thr Thr 1275 Ser Glu Asn Lys 1285 Asp Ser Val Ser Lys 1290 Leu Val Ala Ala 1300 Asp Phe Pro Asn lie 1305 Val Asn Lys 201038734

11151115

Leu lie Val Asn Asp Ser 〇 1205Leu lie Val Asn Asp Ser 〇 1205

Asp Glu Cys Gin Lys Val 1220 149 201038734Asp Glu Cys Gin Lys Val 1220 149 201038734

Val Lys Glu Thr Cys Lys Thr Asp lie Phe lie Glu Leu Gly Ser 1310 1315 1320Val Lys Glu Thr Cys Lys Thr Asp lie Phe lie Glu Leu Gly Ser 1310 1315 1320

Asn Asn Tyr Arg Ser Gly Ala Val Lys Thr lie Leu Gly Pro Glu 1325 1330 1335 工le Val Ser Val Ala 工le Asp Arg Gin Asn Glu Thr Ala Trp Gly 1340 1345 1350Asn Asn Tyr Arg Ser Gly Ala Val Lys Thr lie Leu Gly Pro Glu 1325 1330 1335 work le Val Ser Val Ala work asp Arg Gin Asn Glu Thr Ala Trp Gly 1340 1345 1350

Gin Leu Met Lys Met Val Ala Ser Leu lie Ser His Arg Val Pro 1355 1360 1365Gin Leu Met Lys Met Val Ala Ser Leu lie Ser His Arg Val Pro 1355 1360 1365

Gly Val Glu Leu Lys Lys Leu Tyr His Pro Glu Leu Leu Lys Phe 1370 1375 1380Gly Val Glu Leu Lys Lys Leu Tyr His Pro Glu Leu Leu Lys Phe 1370 1375 1380

Asp Pro Gin Ala Lys Pro Asn Arg Phe lie Arg Asn lie Glu Leu 1385 1390 1395Asp Pro Gin Ala Lys Pro Asn Arg Phe lie Arg Asn lie Glu Leu 1385 1390 1395

Asn Gly Phe Phe Asp Arg Thr Asn lie lie Val Asp Lys Gin Leu 1400 1405 1410Asn Gly Phe Phe Asp Arg Thr Asn lie lie Val Asp Lys Gin Leu 1400 1405 1410

Ser Pro Ala Asp Pro Lys Leu Ala Glu lie Val Asn Asn Arg Asn 1415 1420 1425Ser Pro Ala Asp Pro Lys Leu Ala Glu lie Val Asn Asn Arg Asn 1415 1420 1425

Met Pro Lys Asp Asn Val Tyr Val Pro 工le Glu Arg Val Lys Thr 1430 1435 1440Met Pro Lys Asp Asn Val Tyr Val Pro work le Glu Arg Val Lys Thr 1430 1435 1440

Met lie Lys Ala Glu Pro Ala Asn Leu Gin Val Ser Val Gly Ser 1445 1450 1455Met lie Lys Ala Glu Pro Ala Asn Leu Gin Val Ser Val Gly Ser 1445 1450 1455

Lys Pro Val Val Thr Glu Arg lie Ser Ser Asp Asp Asn Leu Phe 1460 1465 1470Lys Pro Val Val Thr Glu Arg lie Ser Ser Asp Asp Asn Leu Phe 1460 1465 1470

Glu Lys Leu Ser Glu lie Thr Lys Ser Phe Asp Gly Val Asn Ala 1475 1480 1485Glu Lys Leu Ser Glu lie Thr Lys Ser Phe Asp Gly Val Asn Ala 1475 1480 1485

Cys Thr Glu Ala Met Leu Gly Asp Ser Gly Phe Leu Lys Thr Tyr 1490 1495 1500Cys Thr Glu Ala Met Leu Gly Asp Ser Gly Phe Leu Lys Thr Tyr 1490 1495 1500

Glu Val Asp Tyr Pro Leu Tyr Thr Gly Ala Met Ala Lys Gly lie 1505 1510 1515Glu Val Asp Tyr Pro Leu Tyr Thr Gly Ala Met Ala Lys Gly lie 1505 1510 1515

Ala Ser Ala Asp Leu Val lie Ala Ala Gly Lys Ser Lys lie Leu 1520 1525 1530Ala Ser Ala Asp Leu Val lie Ala Ala Gly Lys Ser Lys lie Leu 1520 1525 1530

Ala Ser Phe Gly Ala Gly Gly Leu Ala Leu Gin Val Val Glu Asp 1535 1540 1545Ala Ser Phe Gly Ala Gly Gly Leu Ala Leu Gin Val Val Glu Asp 1535 1540 1545

Ala lie Lys Gin lie Lys Ala Glu Leu Gly Asn Gly Pro Phe Ala 1550 1555 1560Ala lie Lys Gin lie Lys Ala Glu Leu Gly Asn Gly Pro Phe Ala 1550 1555 1560

Val Asn Leu lie His Ser Pro Phe Asp Pro Ser Leu Glu Lys Gly 1565 1570 1575Val Asn Leu lie His Ser Pro Phe Asp Pro Ser Leu Glu Lys Gly 1565 1570 1575

Asn Val Asp Leu Phe Leu Lys Tyr Asn Val Arg Phe Val Glu Val 150 201038734 1580 1585 1590Asn Val Asp Leu Phe Leu Lys Tyr Asn Val Arg Phe Val Glu Val 150 201038734 1580 1585 1590

Ser Ala Phe Met Ser Leu Thr Pro Gin Val Val Arg Tyr Arg Ala 1595 1600 1605Ser Ala Phe Met Ser Leu Thr Pro Gin Val Val Arg Tyr Arg Ala 1595 1600 1605

Ala Gly Leu Ala Lys Ala Arg Asp Gly Ser Val Lys lie Gin Asn 1610 1615 1620Ala Gly Leu Ala Lys Ala Arg Asp Gly Ser Val Lys lie Gin Asn 1610 1615 1620

Arg lie lie Ala Lys lie Ser Arg Thr Glu Leu Ala Glu Leu Phe 1625 1630 1635Arg lie lie Ala Lys lie Ser Arg Thr Glu Leu Ala Glu Leu Phe 1625 1630 1635

Leu Lys Pro Ala Pro Lys Asn lie Leu Asp Ala Leu Val Ala Asp 1640 1645 1650Leu Lys Pro Ala Pro Lys Asn lie Leu Asp Ala Leu Val Ala Asp 1640 1645 1650

Gly Ser lie Ser Gin Glu Gin Ala Gin Leu Ala Leu Leu Val Pro 1655 1660 1665 ΟGly Ser lie Ser Gin Glu Gin Ala Gin Leu Ala Leu Leu Val Pro 1655 1660 1665 Ο

Met Ala Asp Asp lie Thr Val Glu Ala Asp Ser Gly Gly His Thr 1670 1675 1680Met Ala Asp Asp lie Thr Val Glu Ala Asp Ser Gly Gly His Thr 1670 1675 1680

Asp Asn Arg Pro lie His Val Leu Leu Pro Leu lie lie Gin Gin 1685 1690 1695Asp Asn Arg Pro lie His Val Leu Leu Pro Leu lie lie Gin Gin 1685 1690 1695

Arg Asn Arg lie Cys Lys Gin Tyr Pro Lys His Leu Lys Val Arg 1700 1705 1710 lie Gly Ala Ala Gly Gly lie Gly Cys Pro Lys Ala Ala Phe Ala 1715 1720 1725Arg Asn Arg lie Cys Lys Gin Tyr Pro Lys His Leu Lys Val Arg 1700 1705 1710 lie Gly Ala Ala Gly Gly lie Gly Cys Pro Lys Ala Ala Phe Ala 1715 1720 1725

Ala Phe Glu Met Gly Ala Ala Tyr He Ala Thr Gly Thr Val Asn 1730 1735 1740Ala Phe Glu Met Gly Ala Ala Tyr He Ala Thr Gly Thr Val Asn 1730 1735 1740

Gin Leu Ser Lys Glu Ala Gly Thr Cys Asp Tyr Val Arg Lys Val 1745 1750 1755Gin Leu Ser Lys Glu Ala Gly Thr Cys Asp Tyr Val Arg Lys Val 1745 1750 1755

Leu Asn Lys Ala Thr Tyr Ser Asp Val Thr Met Ala Pro Ala Ala 1760 1765 1770Leu Asn Lys Ala Thr Tyr Ser Asp Val Thr Met Ala Pro Ala Ala 1760 1765 1770

Asp Met Phe Asp His Gly Val Glu Leu Gin Val Leu Lys Lys Gly 1775 1780 1785Asp Met Phe Asp His Gly Val Glu Leu Gin Val Leu Lys Lys Gly 1775 1780 1785

Thr Met Phe Pro Ser Arg Ala Lys Lys Leu Tyr Asp Leu Phe Lys 1790 1795 1800Thr Met Phe Pro Ser Arg Ala Lys Lys Leu Tyr Asp Leu Phe Lys 1790 1795 1800

Lys Tyr Lys Ser lie Glu Glu Leu Pro Ala Asp Glu Val Lys Lys 1805 1810 1815Lys Tyr Lys Ser lie Glu Glu Leu Pro Ala Asp Glu Val Lys Lys 1805 1810 1815

Leu Glu Gin Lys Val Phe Lys Lys Ser Phe Asp Glu Val Trp Asp 1820 1825 1830Leu Glu Gin Lys Val Phe Lys Lys Ser Phe Asp Glu Val Trp Asp 1820 1825 1830

Glu Thr Lys Asn Tyr Tyr lie Asn Arg Leu His Ser Pro Glu Lys 1835 1840 1845 lie Glu Arg Ala Glu Arg Asp Ala Lys Leu Lys Met Ser Leu Cys 1850 1855 1860 151 201038734Glu Thr Lys Asn Tyr Tyr lie Asn Arg Leu His Ser Pro Glu Lys 1835 1840 1845 lie Glu Arg Ala Glu Arg Asp Ala Lys Leu Lys Met Ser Leu Cys 1850 1855 1860 151 201038734

Phe Arg Trp Tyr Leu Ser Lys Ser Ser Arg Trp Ala Asn Thr Gly 1865 1870 1875Phe Arg Trp Tyr Leu Ser Lys Ser Ser Arg Trp Ala Asn Thr Gly 1865 1870 1875

Glu Ser Gly Arg Val Gin Asp Tyr Gin lie Trp Cys Gly Pro Ala 1880 1885 1890 lie Gly Ser Tyr Asn Asp Phe Ala Lys Gly Ser Pro Cys Leu Asp 1895 1900 1905Glu Ser Gly Arg Val Gin Asp Tyr Gin lie Trp Cys Gly Pro Ala 1880 1885 1890 lie Gly Ser Tyr Asn Asp Phe Ala Lys Gly Ser Pro Cys Leu Asp 1895 1900 1905

Pro Glu lie Leu Gly Ser Phe Pro Ser Val Val Gin lie Asn Lys 1910 1915 1920Pro Glu lie Leu Gly Ser Phe Pro Ser Val Val Gin lie Asn Lys 1910 1915 1920

His lie Leu Arg Gly Ala Cys Phe Tyr Gin Arg Leu Ser Gin Leu 1925 1930 1935His lie Leu Arg Gly Ala Cys Phe Tyr Gin Arg Leu Ser Gin Leu 1925 1930 1935

Lys Tyr Leu Asn Phe Asn Tyr Glu Glu Leu Asp Thr Leu Thr Tyr 1940 1945 1950Lys Tyr Leu Asn Phe Asn Tyr Glu Glu Leu Asp Thr Leu Thr Tyr 1940 1945 1950

Ser Ala Ser Asn Phe lie 1955 &lt;210&gt; 72 &lt;211&gt; 4368 &lt;212&gt; DNA &lt;213〉 破囊壺菌物種 &lt;400&gt; 72 atggttggtt tacaaatgaa aaagaaacca gtatgggaga tgagtaagga agaacaaagt 60 tctggaaaga atgttgtatt tgactatgat gaattgttgg aatttgctga aggtgatatt 120 ggtaaagtct ttggacctaa gtttgatatt atcgataagt atagtcgacg tgtacgttta 180 cctgcgagag aatatcttct agttaccaga gttactttga tggatgctga agttgggaat 240 ttcagagttg gatctagaat ggttactgaa tatgatgttc cagtaaatgg tgaactttca 300 caaggtggtg atgttccatg ggctgttctt gttgaatctg gacaatgtga tcttatgtta 360 atatcttata tgggtattga ttttcaatgt aaaggtgatc gtgtctatcg attattaaat 420 actacgttga cgttttacgg tgttgctcat gagggtgaaa cactagtata cgatattcgt 480Ser Ala Ser Asn Phe lie 1955 &lt;210&gt; 72 &lt;211&gt; 4368 &lt;212&gt; DNA &lt;213> Thraustochytrium species &lt;400&gt; 72 atggttggtt tacaaatgaa aaagaaacca gtatgggaga tgagtaagga agaacaaagt 60 tctggaaaga atgttgtatt tgactatgat gaattgttgg aatttgctga aggtgatatt 120 ggtaaagtct ttggacctaa gtttgatatt atcgataagt atagtcgacg tgtacgttta 180 cctgcgagag aatatcttct agttaccaga gttactttga tggatgctga agttgggaat 240 ttcagagttg gatctagaat ggttactgaa tatgatgttc cagtaaatgg tgaactttca 300 caaggtggtg atgttccatg ggctgttctt gttgaatctg gacaatgtga tcttatgtta 360 atatcttata tgggtattga ttttcaatgt aaaggtgatc gtgtctatcg attattaaat 420 actacgttga cgttttacgg tgttgctcat gagggtgaaa cactagtata cgatattcgt 480

gtaactggat ttgcaaaagg tatgcacggt gaaatctcca tgtttttttt tgaatatgat 540 tgttatgtga atggacgatt attaatcgaa atgagagatg gttgtgcggg attttttact 600 gatgaagaac ttgcagcagg taaaggagtt attaaaactg ttgctgaact tcataaaaga 660 aaatctattg ttccaaaatc cattaaacct tttgctctaa atccagcagt acacaaaaca 720 atgttttctg aaaatgatat ggaaaaattg tgtgagcgtc aatgggaaaa tgtattgggt 780 agtggacttc aaggtattga ctacaagtta tgtgcacgga aaatgcttat gattgatcgt 840 attactaaaa tacaacataa tggtggtgca tatggtcttg gattattggt tggcgaaaaa 900 attcttgaac gtgatcattg gtattttcca tgccattttg taaaggatca agttatggct 960 ggctcacttg ttagtgatgg ttgcagtcag ctactaaaac tttatatgtt atggttgggt 1020 ttacatgatg tggttccaga ttttcaattt cgtccagttc ctggacaacc aaataaagtt 1080 cgttgccgtg gacaaattag tccacatcgt ggtaaacttg tttatgttat ggaaataaga 1140 gaaatgggat tcaatgaatc aactggacaa ccatatgcta ttgctgatgt tgatattatt 1200 gatgtaaact atgaacttgg tcaatcattt gatatggctg atattgatag ttatggacgt 1260 152 201038734gtaactggat ttgcaaaagg tatgcacggt gaaatctcca tgtttttttt tgaatatgat 540 tgttatgtga atggacgatt attaatcgaa atgagagatg gttgtgcggg attttttact 600 gatgaagaac ttgcagcagg taaaggagtt attaaaactg ttgctgaact tcataaaaga 660 aaatctattg ttccaaaatc cattaaacct tttgctctaa atccagcagt acacaaaaca 720 atgttttctg aaaatgatat ggaaaaattg tgtgagcgtc aatgggaaaa tgtattgggt 780 agtggacttc aaggtattga ctacaagtta tgtgcacgga aaatgcttat gattgatcgt 840 attactaaaa tacaacataa tggtggtgca tatggtcttg gattattggt tggcgaaaaa 900 attcttgaac gtgatcattg gtattttcca tgccattttg taaaggatca agttatggct 960 ggctcacttg ttagtgatgg ttgcagtcag ctactaaaac tttatatgtt atggttgggt 1020 ttacatgatg tggttccaga ttttcaattt cgtccagttc ctggacaacc aaataaagtt 1080 cgttgccgtg gacaaattag tccacatcgt ggtaaacttg tttatgttat ggaaataaga 1140 gaaatgggat tcaatgaatc aactggacaa ccatatgcta ttgctgatgt tgatattatt 1200 gatgtaaact atgaacttgg tcaatcattt gatatggctg atattgatag ttatggacgt 1260 152 201038734

ggtaatttgt caaagaaaat tgtggttgat tttaaaggaa ttgctttgca aatggaaggt 1320 accgtgaaat catcaaatat cattgattct tcaccaaaat caactattat acaaccacct 1380 ccaaattgtc ttcgtggtga tccactggca ccatcacaag ttacatggca tccaatggca 1440 ggagttaatg gggcaccagc tccttcattt agtccatctg attatccacc acgtgctgtt 1500 tgcttcaaac catttcctgg taatccttta gataacgatc atacacctgg taaaatgcct 1560 ttaacatggt ttaatatgtc cgagtttatg tgtggtaaag tatcaaattg tcttggacca 1620 gaatttaaga gatttgataa ctctaaaaca tccagaagtc ctgcctttga tcttgcactt 1680 gttacacgtg ttgtgagtgt atcagatatg gaatttaaac ctcatttaaa tattgatgtt 1740 aatccaagta agggtacaat gataggtgaa tttgattgcc ctgcagatgc gtggtttttt 1800 caaggatcat gtaacgatgg tcatatgccg tattctattg ttatggaaat tgctcttcaa 1860 acttctggtg tattaacttc agttttgaaa gcacctttga ctatggataa agatgatatt 1920 cttttccgca atttggatgc cactgctgaa atggttcgaa gtgatgttga ttgtagaggt 1980 aaaactatca aaaactttac tcaatgtacc ggttacagta tgctcggaaa aatgggaatt 2040 catagattca catttgaatt atctgttgat gatgtagttt tctacaaagg atcaacatct 2100 tttggttggt tcacccctga agtattcgag tcacaagttg gtcttgataa tggtaaaaaa 2160 gtacaaccat ggtatttgga acaaaaatca tctaatgtag taacttatga cgttgcgtcc 2220 actgctggca aggataagtt attttcaaag attggatcta aggatgcaca agttcaaaga 2280 agaaatacac aatgtgagtt tctagatact atgcatatta ttccaaatac tggaaagtac 2340 aacaaaggtt atgctcatgg agaaaagaaa gttaatccaa acgactggtt cttttcctgt 2400 catttctggt ttgatcctgt gatgcctggt tcattaggta ttgaaagtat gtttcaactc 2460 attgaagcat tttcaattga tcaaggaatc gcttcaaaac atggtattgt gaatccaact 2520 tttgctcatt ccaatggaaa aacttcttgg aaatacagag gtcaattgaa taacaaaggt 2580 aaacgaatgg atagtgaaat tcatatcaaa gatattgtca aaaatgctga tggtactgtt 2640 gatttgattg ctgatggatt tttattggtt gattcactaa gagtatactc tgcagatgat 2700 cttcgcgtaa aaattgtacc gggaaccaaa gctgcaccta aatcagtagc tgctgctcca 2760 agacatgttg caacaccaat tccaggagtg ccttcgaata caagcagtgt tgaaatcagt 2820 ttggaatctt tgaagaaaga attgttaaat cttgagaaac cattgtatct tgaaacttcc 2880 aatcatattg taaaacaatt cggtgacgtt aacaatggcc aagcatccgt tattccacca 2940 tgcaccatca atgatttggg tgagcgtagt tttatggaaa catacaatgt tgttgcacca 3000 ctttacactg gagccatggc taaaggtatt gcatctgctg atttggtaat tgcagctggt 3060 aaaagaaaaa ttttgggttc ttttggcgct ggaggcttac caatgcactt ggttcgtgct 3120 tctgttgaaa aaatccaagc cgcacttcca gaaggtccat acgctgtcaa cttgattcat 3180 agtccattcg actcaaatct tgaaaaggga aatgtagatc tatttttgga aaaaggtgtt 3240 catgttgttg aagcatctgc attcactgct ctgaccactc aagtagttcg ttaccgtgca 3300 tgtggtttat ctcgggctaa agacggatct gtattgatca aaaatagaat catcggtaaa 3360 gtttcaagaa ccgaattggc tgaaatgttt ttcagacctg caccacaaaa cttgcttgac 3420 aagcttattg ctagtggaga aatcactaaa gaacaagctt cattggcttt ggaagtacca 3480 153 201038734 atggctgatg atgtagctgt tgaagctgat agcggtggac atactgataa tagaccaatt 3540 catgtaatcc tacctttgat tatcaatcta cgaaatagaa ttcataaaga atgtggtttt 3600 cctgctgctt tgagagttcg cgttggtgct ggtggtggaa ttggttgtcc aagtgctgca 3660 gttgctgcat tcaatatggg agctgcattc ttgattactg gcagcgtcaa ccaagttagc 3720 aaacaatctg gtacgtgtga tatcgttaga aagcaattat ctgaagcttc gtattcagat 3780 attaccatgg caccagcggc tgatatgttt gatcaaggag tcgagcttca agtattaaaa 3840 aaaggaacta tgtttccatc tcgtgcaaag aaattgtatg aattattctg tatgtacaac 3900 tcatttgatg acatgccaaa aagcgaactt caaagactag agaagcgaat ttttcaaaaa 3960 tcgcttgcgg aagtttggga agaaactaaa gatttttata tcaatcgttt gaataatcct 4020 gagaagattg aacatgctga gaagaaagat ccaaagttga agatgtcatt atgctttaga 4080 tggtatttgg gtttaagttc attttgggca aacaatggaa ttaaagaaag atcaatggac 4140 tatcaaattt ggtgtggtcc agcgattggt tcatacaatg attttgtaaa aggaacttat 4200ggtaatttgt caaagaaaat tgtggttgat tttaaaggaa ttgctttgca aatggaaggt 1320 accgtgaaat catcaaatat cattgattct tcaccaaaat caactattat acaaccacct 1380 ccaaattgtc ttcgtggtga tccactggca ccatcacaag ttacatggca tccaatggca 1440 ggagttaatg gggcaccagc tccttcattt agtccatctg attatccacc acgtgctgtt 1500 tgcttcaaac catttcctgg taatccttta gataacgatc atacacctgg taaaatgcct 1560 ttaacatggt ttaatatgtc cgagtttatg tgtggtaaag tatcaaattg tcttggacca 1620 gaatttaaga gatttgataa ctctaaaaca tccagaagtc ctgcctttga tcttgcactt 1680 gttacacgtg ttgtgagtgt atcagatatg gaatttaaac ctcatttaaa tattgatgtt 1740 aatccaagta agggtacaat gataggtgaa tttgattgcc ctgcagatgc gtggtttttt 1800 caaggatcat gtaacgatgg tcatatgccg tattctattg ttatggaaat tgctcttcaa 1860 acttctggtg tattaacttc agttttgaaa gcacctttga ctatggataa agatgatatt 1920 cttttccgca atttggatgc cactgctgaa atggttcgaa gtgatgttga ttgtagaggt 1980 aaaactatca aaaactttac tcaatgtacc ggttacagta tgctcggaaa aatgggaatt 2040 catagattca catttgaatt atctgttgat gatgtagttt tctacaaagg atcaacatc t 2100 tttggttggt tcacccctga agtattcgag tcacaagttg gtcttgataa tggtaaaaaa 2160 gtacaaccat ggtatttgga acaaaaatca tctaatgtag taacttatga cgttgcgtcc 2220 actgctggca aggataagtt attttcaaag attggatcta aggatgcaca agttcaaaga 2280 agaaatacac aatgtgagtt tctagatact atgcatatta ttccaaatac tggaaagtac 2340 aacaaaggtt atgctcatgg agaaaagaaa gttaatccaa acgactggtt cttttcctgt 2400 catttctggt ttgatcctgt gatgcctggt tcattaggta ttgaaagtat gtttcaactc 2460 attgaagcat tttcaattga tcaaggaatc gcttcaaaac atggtattgt gaatccaact 2520 tttgctcatt ccaatggaaa aacttcttgg aaatacagag gtcaattgaa taacaaaggt 2580 aaacgaatgg atagtgaaat tcatatcaaa gatattgtca aaaatgctga tggtactgtt 2640 gatttgattg ctgatggatt tttattggtt gattcactaa gagtatactc tgcagatgat 2700 cttcgcgtaa aaattgtacc gggaaccaaa gctgcaccta aatcagtagc tgctgctcca 2760 agacatgttg caacaccaat tccaggagtg ccttcgaata caagcagtgt tgaaatcagt 2820 ttggaatctt tgaagaaaga attgttaaat cttgagaaac cattgtatct tgaaacttcc 2880 aatcatattg taaaacaatt cggtgacgtt aacaatggcc aagcatccgt t attccacca 2940 tgcaccatca atgatttggg tgagcgtagt catacaatgt tgttgcacca 3000 ctttacactg gagccatggc taaaggtatt gcatctgctg atttggtaat tgcagctggt 3060 aaaagaaaaa ttttgggttc ttttggcgct ggaggcttac caatgcactt ggttcgtgct 3120 tctgttgaaa aaatccaagc cgcacttcca gaaggtccat acgctgtcaa cttgattcat 3180 agtccattcg actcaaatct tgaaaaggga aatgtagatc tatttttgga aaaaggtgtt 3240 catgttgttg aagcatctgc attcactgct ctgaccactc aagtagttcg ttaccgtgca 3300 tgtggtttat ctcgggctaa agacggatct gtattgatca aaaatagaat catcggtaaa tttatggaaa 3360 gtttcaagaa ccgaattggc tgaaatgttt ttcagacctg caccacaaaa cttgcttgac 3420 aagcttattg ctagtggaga aatcactaaa gaacaagctt cattggcttt ggaagtacca 3480 153 201038734 atggctgatg atgtagctgt tgaagctgat agcggtggac atactgataa tagaccaatt 3540 catgtaatcc tacctttgat tatcaatcta cgaaatagaa ttcataaaga atgtggtttt 3600 cctgctgctt tgagagttcg cgttggtgct ggtggtggaa ttggttgtcc aagtgctgca 3660 gttgctgcat tcaatatggg agctgcattc ttgattactg gcagcgtcaa ccaagttagc 3720 aaacaatctg gtacgtgtga tatcgttaga aagcaa ttat ctgaagcttc gtattcagat 3780 attaccatgg caccagcggc tgatatgttt gatcaaggag tcgagcttca agtattaaaa 3840 aaaggaacta tgtttccatc tcgtgcaaag aaattgtatg aattattctg tatgtacaac 3900 tcatttgatg acatgccaaa aagcgaactt caaagactag agaagcgaat ttttcaaaaa 3960 tcgcttgcgg aagtttggga agaaactaaa gatttttata tcaatcgttt gaataatcct 4020 gagaagattg aacatgctga gaagaaagat ccaaagttga agatgtcatt atgctttaga 4080 tggtatttgg gtttaagttc attttgggca aacaatggaa ttaaagaaag atcaatggac 4140 tatcaaattt ggtgtggtcc agcgattggt tcatacaatg Attttgtaaa aggaacttat 4200

ttggatcctg cagtagcagg ttcatatcca tgtgttgttc aaattaacat gcaaattcta 4260 cgcggtgctt gttttcttca acgagttcgt gcaatcaagc acgatccacg attggatatt 4320 gatgtcgatg aagatgtatt tacctatcgt ccagaatcaa ccctatag 4368 &lt;210&gt; 73 &lt;211&gt; 1455 &lt;212&gt; PRT &lt;213〉 破囊壺菌物種 &lt;400&gt; 73ttggatcctg cagtagcagg ttcatatcca tgtgttgttc aaattaacat gcaaattcta 4260 cgcggtgctt gttttcttca acgagttcgt gcaatcaagc acgatccacg attggatatt 4320 gatgtcgatg aagatgtatt tacctatcgt ccagaatcaa ccctatag 4368 &lt; 210 &gt; 73 &lt; 211 &gt; 1455 &lt; 212 &gt; PRT &lt; 213> Thraustochytrium species &lt; 400 &gt; 73

Met Val Gly Leu Gin Met Lys Lys Lys Pro Val Trp Glu Met Ser Lys 15 10 15Met Val Gly Leu Gin Met Lys Lys Lys Pro Val Trp Glu Met Ser Lys 15 10 15

Glu Glu Gin Ser Ser Gly Lys Asn Val Val Phe Asp Tyr Asp Glu Leu 20 25 30Glu Glu Gin Ser Ser Gly Lys Asn Val Val Phe Asp Tyr Asp Glu Leu 20 25 30

Leu Glu Phe Ala Glu Gly Asp lie Gly Lys Val Phe Gly Pro Lys Phe 35 40 45Leu Glu Phe Ala Glu Gly Asp lie Gly Lys Val Phe Gly Pro Lys Phe 35 40 45

Asp lie lie Asp Lys Tyr Ser Arg Arg Val Arg Leu Pro Ala Arg Glu 50 55 60Asp lie lie Asp Lys Tyr Ser Arg Arg Val Arg Leu Pro Ala Arg Glu 50 55 60

Tyr Leu Leu Val Thr Arg Val Thr Leu Met Asp Ala Glu Val Gly Asn 65 70 75 80Tyr Leu Leu Val Thr Arg Val Thr Leu Met Asp Ala Glu Val Gly Asn 65 70 75 80

Phe Arg Val Gly Ser Arg Met Val Thr Glu Tyr Asp Val Pro Val Asn 85 90 95Phe Arg Val Gly Ser Arg Met Val Thr Glu Tyr Asp Val Pro Val Asn 85 90 95

Gly Glu Leu Ser Gin Gly Gly Asp Val Pro Trp Ala Val Leu Val Glu 100 105 110Gly Glu Leu Ser Gin Gly Gly Asp Val Pro Trp Ala Val Leu Val Glu 100 105 110

Ser Gly Gin Cys Asp Leu Met Leu lie Ser Tyr Met Gly lie Asp Phe 115 120 125Ser Gly Gin Cys Asp Leu Met Leu lie Ser Tyr Met Gly lie Asp Phe 115 120 125

Gin Cys Lys Gly Asp Arg Val Tyr Arg Leu Leu Asn Thr Thr Leu Thr 130 135 140Gin Cys Lys Gly Asp Arg Val Tyr Arg Leu Leu Asn Thr Thr Leu Thr 130 135 140

Phe Tyr Gly Val Ala His Glu Gly Glu Thr Leu Val Tyr Asp lie Arg 154 201038734 145 150 155 160 Val Thr Gly Phe Ala Lys Gly Met 165 His Gly Glu lie Ser Met Phe Phe 170 175 Phe Glu Tyr Asp Cys Tyr Val Asn 180 Gly Arg Leu Leu lie Glu Met Arg 185 190 Asp Gly Cys Ala Gly Phe Phe Thr 195 200 Asp Glu Glu Leu Ala Ala Gly Lys 205 Gly Val lie Lys Thr Val Ala Glu 210 215 Leu His Lys Arg Lys Ser 工le Val 220 Pro Lys Ser lie Lys Pro Phe Ala 225 230 Leu Asn Pro Ala Val His Lys Thr 235 240 Met Phe Ser Glu Asn Asp Met Glu O 245 Lys Leu Cys Glu Arg Gin Trp Glu 250 255 Asn Val Leu Gly Ser Gly Leu Gin 260 Gly lie Asp Tyr Lys Leu Cys Ala 265 270 Arg Lys Met Leu Met lie Asp Arg 275 280 lie Thr Lys lie Gin His Asn Gly 285 . Gly Ala Tyr Gly Leu Gly Leu Leu - 290 295 Val Gly Glu Lys lie Leu Glu Arg 300 - Asp His Trp Tyr Phe Pro Cys His ' 305 310 Phe Val Lys Asp Gin Val Met Ala 315 320 Gly Ser Leu Val Ser Asp Gly Cys 325 Ser Gin Leu Leu Lys Leu Tyr Met 330 335 Leu Trp Leu Gly Leu His Asp Val 340 Val Pro Asp Phe Gin Phe Arg Pro 345 350 Val Pro Gly Gin Pro Asn Lys Val 355 360 Arg Cys Arg Gly Gin lie Ser Pro 365 His Arg Gly Lys Leu Val Tyr Val 370 375 Met Glu lie Arg Glu Met Gly Phe 380Phe Tyr Gly Val Ala His Glu Gly Glu Thr Leu Val Tyr Asp lie Arg 154 201038734 145 150 155 160 Val Thr Gly Phe Ala Lys Gly Met 165 His Gly Glu lie Ser Met Phe Phe 170 175 Phe Glu Tyr Asp Cys Tyr Val Asn 180 Gly Arg Leu Leu lie Glu Met Arg 185 190 Asp Gly Cys Ala Gly Phe Phe Thr 195 200 Asp Glu Glu Leu Ala Ala Gly Lys 205 Gly Val lie Lys Thr Val Ala Glu 210 215 Leu His Lys Arg Lys Ser Lys Ser lie Lys Pro Phe Ala 225 230 Leu Asn Pro Ala Val His Lys Thr 235 240 Met Phe Ser Glu Asn Asp Met Glu O 245 Lys Leu Cys Glu Arg Gin Trp Glu 250 255 Asn Val Leu Gly Ser Gly Leu Gin 260 Gly lie Asp Tyr Lys Leu Cys Ala 265 270 Arg Lys Met Leu Met lie Asp Arg 275 280 lie Thr Lys lie Gin His Asn Gly 285 . Gly Ala Tyr Gly Leu Gly Leu Leu - 290 295 Val Gly Glu Lys lie Leu Glu Arg 300 - Asp His Trp Tyr Phe Pro Cys His ' 305 310 Phe Val Lys Asp Gin Val Met Ala 315 320 Gly Ser Leu Val Ser Asp Gly Cys 325 Ser Gin Leu Leu Lys Leu Tyr Met 330 335 Leu Trp Leu Gly Leu His Asp Val 340 Va l Pro Asp Phe Gin Phe Arg Pro 345 350 Val Pro Gly Gin Pro Asn Lys Val 355 360 Arg Cys Arg Gly Gin lie Ser Pro 365 His Arg Gly Lys Leu Val Tyr Val 370 375 Met Glu lie Arg Glu Met Gly Phe 380

Asn Glu Ser Thr Gly Gin Pro Tyr Ala lie Ala Asp Val Asp lie lie 385 390 395 400Asn Glu Ser Thr Gly Gin Pro Tyr Ala lie Ala Asp Val Asp lie lie 385 390 395 400

Asp Val Asn Tyr Glu Leu Gly Gin 405 Ser Phe Asp Met Ala Asp lie Asp 410 415 Ser Tyr Gly Arg Gly Asn Leu Ser 420 Lys Lys lie Val Val Asp Phe Lys 425 430 Gly lie Ala Leu Gin Met Glu Gly 435 440 Thr Val Lys Ser Ser Asn lie lie 445 155 201038734Asp Val Asn Tyr Glu Leu Gly Gin 405 Ser Phe Asp Met Ala Asp lie Asp 410 415 Ser Tyr Gly Arg Gly Asn Leu Ser 420 Lys Lys lie Val Val Asp Phe Lys 425 430 Gly lie Ala Leu Gin Met Glu Gly 435 440 Thr Val Lys Ser Ser Asn lie lie 445 155 201038734

Asp Ser Ser Pro Lys Ser Thr lie 工le Gin Pro Pro Pro Asn Cys Leu 450 455 460Asp Ser Ser Pro Lys Ser Thr lie Le Gin Pro Pro Pro Asn Cys Leu 450 455 460

Arg Gly Asp Pro Leu Ala Pro Ser Gin Val Thr Trp His Pro Met Ala 465 470 475 480Arg Gly Asp Pro Leu Ala Pro Ser Gin Val Thr Trp His Pro Met Ala 465 470 475 480

Gly Val Asn Gly Ala Pro Ala Pro Ser Phe Ser Pro Ser Asp Tyr Pro 485 490 495Gly Val Asn Gly Ala Pro Ala Pro Ser Phe Ser Pro Ser Asp Tyr Pro 485 490 495

Pro Arg Ala Val Cys Phe Lys Pro Phe Pro Gly Asn Pro Leu Asp Asn 500 505 510Pro Arg Ala Val Cys Phe Lys Pro Phe Pro Gly Asn Pro Leu Asp Asn 500 505 510

Asp His Thr Pro Gly Lys Met Pro Leu Thr Trp Phe Asn Met Ser Glu 515 520 525Asp His Thr Pro Gly Lys Met Pro Leu Thr Trp Phe Asn Met Ser Glu 515 520 525

Phe Met Cys Gly Lys Val Ser Asn Cys Leu Gly Pro Glu Phe Lys Arg 530 535 540Phe Met Cys Gly Lys Val Ser Asn Cys Leu Gly Pro Glu Phe Lys Arg 530 535 540

Phe Asp Asn Ser Lys Thr Ser Arg Ser Pro Ala Phe Asp Leu Ala Leu 545 550 555 560Phe Asp Asn Ser Lys Thr Ser Arg Ser Pro Ala Phe Asp Leu Ala Leu 545 550 555 560

Val Thr Arg Val Val Ser Val Ser Asp Met Glu Phe Lys Pro His Leu 565 570 575Val Thr Arg Val Val Ser Val Ser Asp Met Glu Phe Lys Pro His Leu 565 570 575

Asn lie Asp Val Asn Pro Ser Lys Gly Thr Met lie Gly Glu Phe Asp 580 585 590Asn lie Asp Val Asn Pro Ser Lys Gly Thr Met lie Gly Glu Phe Asp 580 585 590

Cys Pro Ala Asp Ala Trp Phe Phe Gin Gly Ser Cys Asn Asp Gly His 595 600 605Cys Pro Ala Asp Ala Trp Phe Phe Gin Gly Ser Cys Asn Asp Gly His 595 600 605

t e Mt e M

r e s r y Tr e s r y T

a V u 1 G t 5 el M6 _—_ a V wy-&lt;—_ G r e s r h T η o 12 G6 u e L a Tx A ea V u 1 G t 5 el M6 ___ a V wy-&lt;-_ G r e s r h T η o 12 G6 u e L a Tx A e

Leu Thr Ser Val Leu Lys Ala Pro Leu Thr Met Asp Lys Asp Asp lie 625 630 635 640Leu Thr Ser Val Leu Lys Ala Pro Leu Thr Met Asp Lys Asp Asp lie 625 630 635 640

Leu Phe Arg Asn Leu Asp Ala Thr Ala Glu Met Val Arg Ser Asp Val 645 650 655Leu Phe Arg Asn Leu Asp Ala Thr Ala Glu Met Val Arg Ser Asp Val 645 650 655

Asp Cys Arg Gly Lys Thr lie Lys Asn Phe Thr Gin Cys Thr Gly Tyr 660 665 670Asp Cys Arg Gly Lys Thr lie Lys Asn Phe Thr Gin Cys Thr Gly Tyr 660 665 670

Ser Met Leu Gly Lys Met Gly lie His Arg Phe Thr Phe Glu Leu Ser 675 680 685Ser Met Leu Gly Lys Met Gly lie His Arg Phe Thr Phe Glu Leu Ser 675 680 685

Val Asp Asp Val Val Phe Tyr Lys Gly Ser Thr Ser Phe Gly Trp Phe 690 695 700Val Asp Asp Val Val Phe Tyr Lys Gly Ser Thr Ser Phe Gly Trp Phe 690 695 700

Thr Pro Glu Val Phe Glu Ser Gin Val Gly Leu Asp Asn Gly Lys Lys 705 710 715 720Thr Pro Glu Val Phe Glu Ser Gin Val Gly Leu Asp Asn Gly Lys Lys 705 710 715 720

Val Gin Pro Trp Tyr Leu Glu Gin Lys Ser Ser Asn Val Val Thr Tyr 725 730 735Val Gin Pro Trp Tyr Leu Glu Gin Lys Ser Ser Asn Val Val Thr Tyr 725 730 735

Asp Val Ala Ser Thr Ala Gly Lys Asp Lys Leu Phe Ser Lys lie Gly 740 745 750 156 201038734Asp Val Ala Ser Thr Ala Gly Lys Asp Lys Leu Phe Ser Lys lie Gly 740 745 750 156 201038734

Ser Lys Asp Ala Gin Val Gin Arg Arg Asn Thr Gin Cys Glu Phe Leu 755 760 765 Asp Thr Met His lie lie Pro Asn Thr Gly Lys Tyr Asn Lys Gly Tyr 770 775 780 Ala 785 His Gly Glu Lys Lys Val Asn Pro Asn Asp Trp Phe Phe Ser Cys 790 795 800 His Phe Trp Phe Asp Pro Val Met Pro Gly Ser Leu Gly lie Glu Ser 805 810 815 Met Phe Gin Leu lie Glu Ala Phe Ser lie Asp Gin Gly lie Ala Ser 820 825 830 Lys His Gly lie Val Asn Pro Thr Phe Ala His Ser Asn Gly Lys Thr 835 840 845 O Ser Trp Lys Tyr Arg Gly Gin Leu Asn Asn Lys Gly Lys Arg Met Asp 850 855 860 Ser 865 Glu lie His 工le Lys Asp lie Val Lys Asn Ala Asp Gly Thr Val 870 875 880 Asp Leu lie Ala Asp Gly Phe Leu Leu Val Asp Ser Leu Arg Val Tyr 885 890 895 - Ser Ala Asp Asp Leu Arg Val Lys lie Val Pro Gly Thr Lys Ala Ala 900 905 910 Pro Lys Ser Val Ala Ala Ala Pro Arg His Val Ala Thr Pro lie Pro 915 920 925Ser Lys Asp Ala Gin Val Gin Arg Arg Asn Thr Gin Cys Glu Phe Leu 755 760 765 Asp Thr Met His lie lie Pro Asn Thr Gly Lys Tyr Asn Lys Gly Tyr 770 775 780 Ala 785 His Gly Glu Lys Lys Val Asn Pro Asn Asp Trp Phe Phe Ser Cys 790 795 800 His Phe Trp Phe Asp Pro Val Met Pro Gly Ser Leu Gly lie Glu Ser 805 810 815 Met Phe Gin Leu lie Glu Ala Phe Ser lie Asp Gin Gly lie Ala Ser 820 825 830 Lys His Gly lie Val Asn Pro Thr Phe Ala His Ser Asn Gly Lys Thr 835 840 845 O Ser Trp Lys Tyr Arg Gly Gin Leu Asn Asn Lys Gly Lys Arg Met Asp 850 855 860 Ser 865 Glu lie His work Ly Lys Asp lie Val Lys Asn Ala Asp Gly Thr Val 870 875 880 Asp Leu lie Ala Asp Gly Phe Leu Leu Val Asp Ser Leu Arg Val Tyr 885 890 895 - Ser Ala Asp Asp Leu Arg Val Lys lie Val Pro Gly Thr Lys Ala Ala 900 905 910 Pro Lys Ser Val Ala Ala Ala Pro Arg His Val Ala Thr Pro lie Pro 915 920 925

Gly Val Pro Ser Asn Thr Ser Ser Val Glu lie Ser Leu Glu Ser Leu 930 935 940Gly Val Pro Ser Asn Thr Ser Ser Val Glu lie Ser Leu Glu Ser Leu 930 935 940

Lys ❹ 945 Lys Glu Leu Leu Asn Leu Glu Lys Pro Leu Tyr Leu Glu Thr Ser 950 955 960 Asn His lie Val Lys Gin Phe Gly Asp Val Asn Asn Gly Gin Ala Ser 965 970 975 Val lie Pro Pro Cys Thr lie Asn Asp Leu Gly Glu Arg Ser Phe Met 980 985 990 Glu Thr Tyr Asn Val Val Ala Pro Leu Tyr Thr Gly Ala Met Ala Lys 995 1000 1005Lys 945 945 Lys Glu Leu Leu Asn Leu Glu Lys Pro Leu Tyr Leu Glu Thr Ser 950 955 960 Asn His lie Val Lys Gin Phe Gly Asp Val Asn Asn Gly Gin Ala Ser 965 970 975 Val lie Pro Pro Cys Thr lie Asn Asp Leu Gly Glu Arg Ser Phe Met 980 985 990 Glu Thr Tyr Asn Val Val Ala Pro Leu Tyr Thr Gly Ala Met Ala Lys 995 1000 1005

Gly lie Ala Ser Ala Asp Leu Val lie Ala Ala Gly Lys Arg Lys 1010 1015 1020 lie Leu Gly Ser Phe Gly Ala Gly Gly Leu Pro Met His Leu Val 1025 1030 1035 Arg Ala Ser Val Glu Lys lie Gin Ala Ala Leu Pro Glu Gly Pro 1040 1045 1050 157 201038734Gly lie Ala Ser Ala Asp Leu Val lie Ala Ala Gly Lys Arg Lys 1010 1015 1020 lie Leu Gly Ser Phe Gly Ala Gly Gly Leu Pro Met His Leu Val 1025 1030 1035 Arg Ala Ser Val Glu Lys lie Gin Ala Ala Leu Pro Glu Gly Pro 1040 1045 1050 157 201038734

Tyr Ala Val Asn Leu lie His Ser Pro Phe Asp Ser Asn Leu Glu 1055 1060 1065Tyr Ala Val Asn Leu lie His Ser Pro Phe Asp Ser Asn Leu Glu 1055 1060 1065

Lys Gly Asn Val Asp Leu Phe Leu Glu Lys Gly Val His Val Val 1070 1075 1080Lys Gly Asn Val Asp Leu Phe Leu Glu Lys Gly Val His Val Val 1070 1075 1080

Glu Ala Ser Ala Phe Thr Ala Leu Thr Thr Gin Val Val Arg Tyr 1085 1090 1095Glu Ala Ser Ala Phe Thr Ala Leu Thr Thr Gin Val Val Arg Tyr 1085 1090 1095

Arg Ala Cys Gly Leu Ser Arg Ala Lys Asp Gly Ser Val Leu lie 1100 1105 1110Arg Ala Cys Gly Leu Ser Arg Ala Lys Asp Gly Ser Val Leu lie 1100 1105 1110

Lys Asn Arg lie lie Gly Lys Val Ser Arg Thr Glu Leu Ala Glu 1115 1120 1125Lys Asn Arg lie lie Gly Lys Val Ser Arg Thr Glu Leu Ala Glu 1115 1120 1125

Met Phe Phe Arg Pro Ala Pro Gin Asn Leu Leu Asp Lys Leu lie 1130 1135 1140Met Phe Phe Arg Pro Ala Pro Gin Asn Leu Leu Asp Lys Leu lie 1130 1135 1140

Ala Ser Gly Glu lie Thr Lys Glu Gin Ala Ser Leu Ala Leu Glu 1145 1150 1155Ala Ser Gly Glu lie Thr Lys Glu Gin Ala Ser Leu Ala Leu Glu 1145 1150 1155

Val Pro Met Ala Asp Asp Val Ala Val Glu Ala Asp Ser Gly Gly 1160 1165 1170Val Pro Met Ala Asp Asp Val Ala Val Glu Ala Asp Ser Gly Gly 1160 1165 1170

His Thr Asp Asn Arg Pro lie His Val lie Leu Pro Leu lie lie 1175 1180 1185His Thr Asp Asn Arg Pro lie His Val lie Leu Pro Leu lie lie 1175 1180 1185

Asn Leu Arg Asn Arg lie His Lys Glu Cys Gly Phe Pro Ala Ala 1190 1195 1200Asn Leu Arg Asn Arg lie His Lys Glu Cys Gly Phe Pro Ala Ala 1190 1195 1200

Leu Arg Val Arg Val Gly Ala Gly Gly Gly lie Gly Cys Pro Ser 1205 1210 1215Leu Arg Val Arg Val Gly Ala Gly Gly Gly lie Gly Cys Pro Ser 1205 1210 1215

Ala Ala Val Ala Ala Phe Asn Met Gly Ala Ala Phe Leu lie Thr 1220 1225 1230 iAla Ala Val Ala Ala Phe Asn Met Gly Ala Ala Phe Leu lie Thr 1220 1225 1230 i

Gly Ser Val Asn Gin Val Ser Lys Gin Ser Gly Thr Cys Asp lie 1235 1240 1245Gly Ser Val Asn Gin Val Ser Lys Gin Ser Gly Thr Cys Asp lie 1235 1240 1245

Val Arg Lys Gin Leu Ser Glu Ala Ser Tyr Ser Asp lie Thr Met 1250 1255 1260Val Arg Lys Gin Leu Ser Glu Ala Ser Tyr Ser Asp lie Thr Met 1250 1255 1260

Ala Pro Ala Ala Asp Met Phe Asp Gin Gly Val Glu Leu Gin Val 1265 1270 1275Ala Pro Ala Ala Asp Met Phe Asp Gin Gly Val Glu Leu Gin Val 1265 1270 1275

Leu Lys Lys Gly Thr Met Phe Pro Ser Arg Ala Lys Lys Leu Tyr 1280 1285 1290Leu Lys Lys Gly Thr Met Phe Pro Ser Arg Ala Lys Lys Leu Tyr 1280 1285 1290

Glu Leu Phe Cys Met Tyr Asn Ser Phe Asp Asp Met Pro Lys Ser 1295 1300 1305Glu Leu Phe Cys Met Tyr Asn Ser Phe Asp Asp Met Pro Lys Ser 1295 1300 1305

Glu Leu Gin Arg Leu Glu Lys Arg lie Phe Gin Lys Ser Leu Ala 1310 1315 1320Glu Leu Gin Arg Leu Glu Lys Arg lie Phe Gin Lys Ser Leu Ala 1310 1315 1320

Glu Val Trp Glu Glu Thr Lys Asp Phe Tyr lie Asn Arg Leu Asn 158 201038734 1325 1330 1335Glu Val Trp Glu Glu Thr Lys Asp Phe Tyr lie Asn Arg Leu Asn 158 201038734 1325 1330 1335

Asn Pro 1340Asn Pro 1340

Glu Lys lie Glu His 1345Glu Lys lie Glu His 1345

Ala Glu Lys Lys Asp 1350Ala Glu Lys Lys Asp 1350

Lys Met 1355Lys Met 1355

Ser Leu Cys Phe Arg 1360Ser Leu Cys Phe Arg 1360

Trp Tyr Leu Gly Leu 1365Trp Tyr Leu Gly Leu 1365

Pro Lys Leu Ser Ser PhePro Lys Leu Ser Ser Phe

Trp Ala 1370Trp Ala 1370

Asn Asn Gly lie Lys 1375Asn Asn Gly lie Lys 1375

Glu Arg Ser Met Asp 1380Glu Arg Ser Met Asp 1380

Trp Cys Gly Pro Ala lie Gly 1385 1390Trp Cys Gly Pro Ala lie Gly 1385 1390

Ser Tyr Asn Asp Phe 1395Ser Tyr Asn Asp Phe 1395

Thr Tyr Leu Asp Pro Ala Val 1400 1405Thr Tyr Leu Asp Pro Ala Val 1400 1405

Ala Gly Ser Tyr Pro 1410Ala Gly Ser Tyr Pro 1410

Gin lie Asn Met Gin lie Leu 1415 1420Gin lie Asn Met Gin lie Leu 1415 1420

Arg Gly Ala Cys Phe 1425Arg Gly Ala Cys Phe 1425

Tyr Gin lie Val Lys Gly Cys Val Val Leu Gin ArgTyr Gin lie Val Lys Gly Cys Val Val Leu Gin Arg

Val Arg Ala lie Lys His Asp 1430 1435Val Arg Ala lie Lys His Asp 1430 1435

Pro Arg Leu Asp lie Asp Val Asp 1440Pro Arg Leu Asp lie Asp Val Asp 1440

Glu Asp Val Phe Thr Tyr Arg Pro Glu Ser Thr Leu 1445 1450 1455Glu Asp Val Phe Thr Tyr Arg Pro Glu Ser Thr Leu 1445 1450 1455

&lt;210&gt; 74 &lt;211&gt; 1350 &lt;212&gt; DNA &lt;213〉 破囊壺菌物種 &lt;400&gt; 74 agaattgcta tgggaagcaa gttactgcgt gggtttattc gaagattctg gctaatatac ggtggtcaaa gttttaagaa gctaattttc ggccgttgta gctagttctt gcaatgattg aaaacacctg atgcttattg gatggtgata tctggtattt agagctggtg ttgttggatt tacgtgatgg attataatcc ctgaatatga atgctaatca ctgcatttac aagcatctca aaatgggatt ctgaatggag ctaatacatt taattgctat ctggtgcaac ttttttcaac gtgaaggttc ctgtacatgc atacaccaac tatcaccaaa atctgcgatt tttgaattgt aacaaaaggt atttgattct aacgttaact taatgagaaa tgaattttat acctgatgag attagattcc caatatggaa taaagttgct ttgtactgat tgatcaaagt agctatgttt tgttatacgt tatttctggt tactattact ttaccaagtg ttaagtgatt gtaaaggata agagaatttg ttattaaagg aaaaatattg tcaagactta gatgttgaaa tttcctggtt ggtatgaatt attgatgaat aacgctcttg tgtcttgcat gttttaaaac tcatgttcat caagaagaag ttagttgaag gtgaaaatgt tacctgcgga aaatttattg gacttaatat ttaaagaagc gttgtgttct attatgttgt ctgctgttga ttcttggtaa gtgttgtaga tattacatgg gtatgtatat atgatgaaaa gttatgctga catcatctga ctattcttag gacatggtac tagagaatct tcgtgttgat taaacgtggt gttacaaatg attagatgat tggtattggt tgtggataaa aaagtttaaa tgttactgct tgctgcttgt tgattgtgat ggcattttca aacaaaaggt cgcagtgaga cggtaaagca agcatatcgt tggtacacca 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 159 201038734 gtgggtgata aaattgaatt aacagcttta cgcaatgtat ttgataaagc atatggtcct 1080 ggtcataagg aagaagttgc tgttggaagt attaaaagtc aaattggtca tttgaaagct 1140 gttgctggtt gtgctggtct tgtgaaattg gttatggcat tgaaacataa aacactacct 1200 caaagtatta atgttgaaaa tccacctaat ttagtggatg gtactgtcat tagtgatact 1260 actttatata ttaatacaat gaatcgtcca tggattacta agcctggtgt tccaagaaga 1320 gctggtatat ctagtttcgg atttggtggt 1350 &lt;210&gt; 75 &lt;211&gt; 450 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 75&Lt; 210 &gt; 74 &lt; 211 &gt; 1350 &lt; 212 &gt; DNA &lt; 213> Thraustochytrium species &lt; 400 &gt; 74 agaattgcta tgggaagcaa gttactgcgt gggtttattc gaagattctg gctaatatac ggtggtcaaa gttttaagaa gctaattttc ggccgttgta gctagttctt gcaatgattg aaaacacctg atgcttattg gatggtgata tctggtattt agagctggtg ttgttggatt tacgtgatgg attataatcc ctgaatatga atgctaatca ctgcatttac aagcatctca aaatgggatt ctgaatggag ctaatacatt taattgctat ctggtgcaac ttttttcaac gtgaaggttc ctgtacatgc atacaccaac tatcaccaaa atctgcgatt tttgaattgt aacaaaaggt atttgattct aacgttaact taatgagaaa tgaattttat acctgatgag attagattcc caatatggaa taaagttgct ttgtactgat tgatcaaagt agctatgttt tgttatacgt tatttctggt tactattact ttaccaagtg ttaagtgatt gtaaaggata agagaatttg ttattaaagg aaaaatattg tcaagactta gatgttgaaa tttcctggtt ggtatgaatt attgatgaat aacgctcttg tgtcttgcat gttttaaaac tcatgttcat caagaagaag ttagttgaag gtgaaaatgt tacctgcgga aaatttattg Gacttaatat ttaaagaagc gttgtgttct attatgttgt ctgctgttga ttcttggtaa gtgttgtaga tattacatgg gtatgtatat atgatgaaaa gttatgctga catca tctga ctattcttag gacatggtac tagagaatct tcgtgttgat taaacgtggt gttacaaatg attagatgat tggtattggt tgtggataaa aaagtttaaa tgttactgct tgctgcttgt tgattgtgat ggcattttca aacaaaaggt cgcagtgaga cggtaaagca agcatatcgt tggtacacca 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 159 201038734 gtgggtgata 1080 ggtcataagg aagaagttgc aaattgaatt aacagcttta cgcaatgtat ttgataaagc atatggtcct tgttggaagt attaaaagtc aaattggtca tttgaaagct 1140 gttgctggtt gtgctggtct tgtgaaattg gttatggcat tgaaacataa aacactacct 1200 caaagtatta atgttgaaaa tccacctaat ttagtggatg gtactgtcat tagtgatact 1260 actttatata ttaatacaat gaatcgtcca tggattacta agcctggtgt tccaagaaga 1320 gctggtatat ctagtttcgg atttggtggt 1350 &lt; 210 &gt; 75 &lt; 211 &gt; 450 &lt; 212 &gt; PRT &lt; 213 &gt; Thraustochytrium Bacterial species &lt;400&gt; 75

Arg 工le Ala lie Val Gly Leu Ser Ala lie Leu Pro Ser Gly Glu Asn 15 10 15Arg work le Ala lie Val Gly Leu Ser Ala lie Leu Pro Ser Gly Glu Asn 15 10 15

Val Arg Glu Ser Trp Glu Ala lie Arg Asp Gly Leu Asn Cys Leu Ser 20 25 30Val Arg Glu Ser Trp Glu Ala lie Arg Asp Gly Leu Asn Cys Leu Ser 20 25 30

Asp Leu Pro Ala Asp Arg Val Asp Val Thr Ala Tyr Tyr Asn Pro Thr 35 40 45Asp Leu Pro Ala Asp Arg Val Asp Val Thr Ala Tyr Tyr Asn Pro Thr 35 40 45

Lys Gly Val Lys Asp Lys lie Tyr Cys Lys Arg Gly Gly Phe lie Pro 50 55 60Lys Gly Val Lys Asp Lys lie Tyr Cys Lys Arg Gly Gly Phe lie Pro 50 55 60

Glu Tyr Glu Phe Asp Ser Arg Glu Phe Gly Leu Asn Met Leu Gin Met 65 70 75 80Glu Tyr Glu Phe Asp Ser Arg Glu Phe Gly Leu Asn Met Leu Gin Met 65 70 75 80

Glu Asp Ser Asp Ala Asn Gin Thr Leu Thr Leu Leu Lys Val Lys Glu 85 90 95Glu Asp Ser Asp Ala Asn Gin Thr Leu Thr Leu Leu Lys Val Lys Glu 85 90 95

Ala Leu Asp Asp Ala Asn lie Pro Ala Phe Thr Asn Glu Lys Lys Asn 100 105 110 lie Gly Cys Val Leu Gly lie Gly Gly Gly Gin Lys Ala Ser His Glu 115 120 125Ala Leu Asp Asp Ala Asn lie Pro Ala Phe Thr Asn Glu Lys Lys Asn 100 105 110 lie Gly Cys Val Leu Gly lie Gly Gly Gly Gin Lys Ala Ser His Glu 115 120 125

Phe Tyr Ser Arg Leu Asn Tyr Val Val Val Asp Lys Val Leu Arg Lys 130 135 140Phe Tyr Ser Arg Leu Asn Tyr Val Val Val Asp Lys Val Leu Arg Lys 130 135 140

Met Gly Leu Pro Asp Glu Asp Val Glu Thr Ala Val Glu Lys Phe Lys 145 150 155 160Met Gly Leu Pro Asp Glu Asp Val Glu Thr Ala Val Glu Lys Phe Lys 145 150 155 160

Ala Asn Phe Pro Glu Trp Arg Leu Asp Ser Phe Pro Gly Phe Leu Gly 165 170 175Ala Asn Phe Pro Glu Trp Arg Leu Asp Ser Phe Pro Gly Phe Leu Gly 165 170 175

Asn Val Thr Ala Gly Arg Cys Thr Asn Thr Phe Asn Met Glu Gly Met 180 185 190Asn Val Thr Ala Gly Arg Cys Thr Asn Thr Phe Asn Met Glu Gly Met 180 185 190

Asn Cys Val Val Asp Ala Ala Cys Ala Ser Ser Leu lie Ala lie Lys 195 200 205Asn Cys Val Val Asp Ala Ala Cys Ala Ser Ser Leu lie Ala lie Lys 195 200 205

Val Ala lie Asp Glu Leu Leu His Gly Asp Cys Asp Ala Met lie Ala 210 215 220 160 201038734Val Ala lie Asp Glu Leu Leu His Gly Asp Cys Asp Ala Met lie Ala 210 215 220 160 201038734

Gly Ala Thr Cys Thr Asp Asn Ala Leu Gly Met Tyr Met Ala Phe Ser 225 230 235 240Gly Ala Thr Cys Thr Asp Asn Ala Leu Gly Met Tyr Met Ala Phe Ser 225 230 235 240

Lys Thr Pro Val Phe Ser Thr Asp Gin Ser Cys Leu Ala Tyr Asp Glu 245 250 255Lys Thr Pro Val Phe Ser Thr Asp Gin Ser Cys Leu Ala Tyr Asp Glu 245 250 255

Lys Thr Lys Gly Met Leu He Gly Glu Gly Ser Ala Met Phe Val Leu 260 265 270Lys Thr Lys Gly Met Leu He Gly Glu Gly Ser Ala Met Phe Val Leu 260 265 270

Lys Arg Tyr Ala Asp Ala Val Arg Asp Gly Asp Thr Val His Ala Val 275 280 285 lie Arg Ser Cys Ser Ser Ser Ser Asp Gly Lys Ala Ser Glv lie Tvr 290 295 300Lys Arg Tyr Ala Asp Ala Val Arg Asp Gly Asp Thr Val His Ala Val 275 280 285 lie Arg Ser Cys Ser Ser Ser Ser Asp Gly Lys Ala Ser Glv lie Tvr 290 295 300

Thr Pro Thr lie Ser Gly Gin Glu Glu Ala lie Leu Arg Ala Tyr Arg 305 310 315 320Thr Pro Thr lie Ser Gly Gin Glu Glu Ala lie Leu Arg Ala Tyr Arg 305 310 315 320

OO

a V y _—I G a _—_ A g r A r 5 e 2 s 3a V y _—I G a _—_ A g r A r 5 e 2 s 3

Pro Asn Thr lie Thr Leu Val Glu Gly His Gly 330 335Pro Asn Thr lie Thr Leu Val Glu Gly His Gly 330 335

Thr Gly Thr Pro Val Gly Asp Lys lie Glu Leu Thr Ala Leu Arg Asn 340 345 350Thr Gly Thr Pro Val Gly Asp Lys lie Glu Leu Thr Ala Leu Arg Asn 340 345 350

Val Phe Asp Lys Ala Tyr Gly Pro Gly His Lys Glu Glu Val Ala Val 355 360 365Val Phe Asp Lys Ala Tyr Gly Pro Gly His Lys Glu Glu Val Ala Val 355 360 365

Gly Ser lie Lys Ser Gin lie Gly His Leu Lys Ala Val Ala Gly Cys 370 375 380Gly Ser lie Lys Ser Gin lie Gly His Leu Lys Ala Val Ala Gly Cys 370 375 380

Ala Gly Leu Val Lys Leu Val Met Ala Leu Lys His Lys Thr Leu Pro 385 390 395 400Ala Gly Leu Val Lys Leu Val Met Ala Leu Lys His Lys Thr Leu Pro 385 390 395 400

r e s n .—I Gr e s n .—I G

Tx L〇 a o V4 n s ATx L〇 a o V4 n s A

Glu Asn Pro Pro Asn Leu Val Asp Gly Thr Val 410 415 lie Ser Asp Thr Thr Leu Tyr lie Asn Thr Met Asn Arg Pro Trp lie 420 425 430Glu Asn Pro Pro Asn Leu Val Asp Gly Thr Val 410 415 lie Ser Asp Thr Thr Leu Tyr lie Asn Thr Met Asn Arg Pro Trp lie 420 425 430

Thr Lys Pro Gly Val Pro Arg Arg Ala Gly lie Ser Ser Phe Gly Phe 435 440 445Thr Lys Pro Gly Val Pro Arg Arg Ala Gly lie Ser Ser Phe Gly Phe 435 440 445

Gly Gly 450 &lt;210&gt; 76 &lt;211&gt; 921 &lt;212&gt; DNA &lt;213&gt;破囊壺菌物種 &lt;400&gt; 76 ttttctggac aaggcgcaca atatacccat atgtttaatg atgttgcaat gcaatggcca 60 caatttcgtt tatgtgtaaa tgatatggag aaagcacagg aagaagttat caatgataaa 120 agtgtgaaac gtatcagtca agttatgttt cctcgtaaac catatgcaag agaatcacct 180 ttagacaata aagaaatctc taagactgaa tattctcaaa caacaactgt cgctagttca 240 161 201038734 gtaggtttat tctttaggtg aagttggtat gcaatggctg tgggttgcta caagctgaaa ggggcatttc gcagttaagt ggtgtattta cttactcaaa aaacaagtac gtttcggtga ttgaaatttt aatttagtgc gtaatcgtgc ctgttattgg acaataactc caagtaaatt attcgcctca ttaataaacc cggatccaaa ttaagaatat tttccaaatt atccagctag ccgtgatgct attgtatgca aatggctatg tccaaatgct tccatctcaa gaaaactcaa tatggaaaat aactggttct aactgctttg gtacgcggct ggtcaatgaa t ggtttcgctc gctggattga agagatgcac tcttcaatta actgttatta ggtttccgtg gctgaaaagc tctccaaaaa tcaagacata ggagctcgtg atttttcctg ctgcttttgt ttgatcgcga caaaaaaatc agctttcagc ccggtgcaaa tggttcattt aatttcaaaa ttttcagcaa tgactagttc tctttattga gtgatacaag tgctggtcat agatttattc tgctgatgga tcctgaagta ttctggtgta ggcatgtgat agctctttca tgtaactggt tgtacaattt atttggacca cgttttaact 300 360 420 480 540 600 660 720 780 840 900 921 &lt;210&gt; 77 &lt;211&gt; 307 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 77 Phe Ser Gly Gin Gly Ala Gin Tyr Thr His Met Phe Asn Asp Val Ala 15 10 15 Met Gin Trp Pro Gin Phe Arg Leu Cys Val Asn Asp Met Glu Lys Ala 20 25 30 Gin Glu Glu Val lie Asn Asp Lys Ser Val Lys Arg lie Ser Gin Val 35 40 45 Met Phe Pro Arg Lys Pro Tyr Ala Arg Glu Ser Pro Leu Asp Asn Lys 50 55 60 oGly Gly 450 &lt;210&gt; 76 &lt;211&gt; 921 &lt;212&gt; DNA &lt;213&gt; Thraustochytrium species &lt;400&gt; 76 ttttctggac aaggcgcaca atatacccat atgtttaatg atgttgcaat gcaatggcca 60 caatttcgtt tatgtgtaaa tgatatggag aaagcacagg aagaagttat caatgataaa 120 agtgtgaaac gtatcagtca agttatgttt cctcgtaaac catatgcaag agaatcacct 180 ttagacaata aagaaatctc taagactgaa tattctcaaa caacaactgt cgctagttca 240 161 201038734 gtaggtttat tctttaggtg aagttggtat gcaatggctg tgggttgcta caagctgaaa ggggcatttc gcagttaagt ggtgtattta cttactcaaa aaacaagtac gtttcggtga ttgaaatttt aatttagtgc gtaatcgtgc ctgttattgg acaataactc caagtaaatt attcgcctca ttaataaacc cggatccaaa ttaagaatat tttccaaatt atccagctag ccgtgatgct attgtatgca aatggctatg tccaaatgct tccatctcaa gaaaactcaa tatggaaaat aactggttct aactgctttg gtacgcggct ggtcaatgaa t ggtttcgctc gctggattga Agagatgcac tcttcaatta actgttatta ggtttccgtg gctgaaaagc tctccaaaaa tcaagacata ggagctcgtg atttttcctg ctgcttttgt ttgatcgcga caaaaaaatc agctttcagc ccggtgcaaa tggttcattt aatttcaaaa ttttcagcaa tgactag Ttc tctttattga gtgatacaag tgctggtcat agatttattc tgctgatgga tcctgaagta ttctggtgta ggcatgtgat agctctttca tgtaactggt tgtacaattt atttggacca cgttttaact 300 360 420 480 540 600 660 720 780 840 900 921 &lt;210&gt; 77 &lt;211&gt; 307 &lt;212&gt; PRT &lt;213&gt; Thraustochytrium species &lt;400&gt; 77 Phe Ser Gly Gin Gly Ala Gin Tyr Thr His Met Phe Asn Asp Val Ala 15 10 15 Met Gin Trp Pro Gin Phe Arg Leu Cys Val Asn Asp Met Glu Lys Ala 20 25 30 Gin Glu Glu Val lie Asn Asp Lys Ser Val Lys Arg lie Ser Gin Val 35 40 45 Met Phe Pro Arg Lys Pro Tyr Ala Arg Glu Ser Pro Leu Asp Asn Lys 50 55 60 o

Glu lie Ser Lys Thr Glu Tyr Ser Gin Thr Thr Thr Val Ala Ser Ser 65 70 75 80 Val Gly Leu Phe Glu lie Phe Arg Asp Ala Gly Phe Ala Pro Ala Phe 85 90 95 1Glu lie Ser Lys Thr Glu Tyr Ser Gin Thr Thr Thr Val Ala Ser Ser 65 70 75 80 Val Gly Leu Phe Glu lie Phe Arg Asp Ala Gly Phe Ala Pro Ala Phe 85 90 95 1

Val Ala Gly His Ser Leu Gly Glu Phe Ser Ala Leu Tyr Ala Ala Gly 100 105 110 Leu lie Asp Arg Glu Asp Leu Phe Lys Leu Val Cys Asn Arg Ala Met 115 120 125Val Ala Gly His Ser Leu Gly Glu Phe Ser Ala Leu Tyr Ala Ala Gly 100 105 110 Leu lie Asp Arg Glu Asp Leu Phe Lys Leu Val Cys Asn Arg Ala Met 115 120 125

Ala Met Arg Asp Ala Pro Lys Lys Ser Ala Asp Gly Ala Met Ala Ala 130 135 140 1 5 a 4 V1 lie Gly Pro Asn Ala Ser Ser lie Lys Leu Ser Ala Pro Glu Val 150 155 160Ala Met Arg Asp Ala Pro Lys Lys Ser Ala Asp Gly Ala Met Ala Ala 130 135 140 1 5 a 4 V1 lie Gly Pro Asn Ala Ser Ser lie Lys Leu Ser Ala Pro Glu Val 150 155 160

Trp Val Ala Asn Asn Asn Ser Pro Ser Gin Thr Val lie Thr Gly Ala 165 170 175 162 201038734Trp Val Ala Asn Asn Asn Ser Pro Ser Gin Thr Val lie Thr Gly Ala 165 170 175 162 201038734

Asn Ser Gly Val Gin Ala Glu Thr Ser Lys Leu Lys Thr Gin Gly Phe 180 185 190Asn Ser Gly Val Gin Ala Glu Thr Ser Lys Leu Lys Thr Gin Gly Phe 180 185 190

Arg Val Val His Leu Ala Cys Asp Gly Ala Phe His Ser Pro His Met 195 200 205Arg Val Val His Leu Ala Cys Asp Gly Ala Phe His Ser Pro His Met 195 200 205

Glu Asn Ala Glu Lys Gin Phe Gin Lys Ala Leu 210 215 A r o e 2 S2 vaGlu Asn Ala Glu Lys Gin Phe Gin Lys Ala Leu 210 215 A r o e 2 S2 va

Ly e h pLy e h p

Asn Lys Pro Thr Gly Ser Ser Pro Lys lie Phe Ser Asn Val Thr Gly 225 230 235 240Asn Lys Pro Thr Gly Ser Ser Pro Lys lie Phe Ser Asn Val Thr Gly 225 230 235 240

Gly Val Phe Thr Asp Pro Lys Thr Ala Leu Ser Arg His Met Thr Ser 245 250 255Gly Val Phe Thr Asp Pro Lys Thr Ala Leu Ser Arg His Met Thr Ser 245 250 255

Ser Val Gin Phe Leu Thr Gin lie Lys Asn Met Tyr Ala Ala Gly Ala 260 265 270 o 〇Ser Val Gin Phe Leu Thr Gin lie Lys Asn Met Tyr Ala Ala Gly Ala 260 265 270 o 〇

Arg Val Phe lie Glu Phe Gly Pro Lys Gin Val Leu Ser Lys Leu Val 275 280 285Arg Val Phe lie Glu Phe Gly Pro Lys Gin Val Leu Ser Lys Leu Val 275 280 285

Asn Glu lie Phe Pro Gly Asp Thr Ser Val Leu Thr Val Ser Val Asn 290 295 300Asn Glu lie Phe Pro Gly Asp Thr Ser Val Leu Thr Val Ser Val Asn 290 295 300

Pro Ala Ser 305 &lt;210&gt; 78 &lt;211&gt; 3303 &lt;212&gt; DNA &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 78 gcaagtccag ctaccgttcg tgtcgtttca gctcctgttc aagcggctgc tcctgtgcag 60 gtatctgctt ctgttgattc tggtttgttg gcaaaagcgg aacaagttgt attggaagta 120 ttggcatcga agactggtta tgagactgag ttgattgaat tggatatgga attggaaact 180 gaacttggta ttgattctat caagagagta gaaattcttt ctgaagttca agctcaattg 240 aatgttgaag ctaaagatgt agatgctctt agtagaactc gtactgttgg tgaagtgatt 300 gatgcaatga aagccgaaat tgctggtggt caaccagctg ctcctgttca agttgcagct 360 cctactcaag tagttgctcc tgttcaagca tctgctcctg ttgattctgg tttgttagca 420 aaagcggaac aagttgtatt ggaagtattg gcatcgaaga ctggttatga gactgagttg 480 attgaattgg atatggaatt ggaaaccgaa cttggtattg attctatcaa gagagtagaa 540 attctttctg aagttcaagc tcaattgagt gttgaagcta aagatgtaga tgctcttagt 600 agaactcgta ctgttggtga agtgattgat gcaatgaaag ccgaaattgc tggtggtcaa 660 ccagctgctc ctgttcaagt tgcagctcct actcaagtag ttgctcctgt tcaagcatct 720 gctcctgttg attctggttt gttagcaaaa gcggaacaag ttgtattgga agtattggca 780 tcgaagactg gttatgagac tgagttgatt gaattggata tggaattgga aaccgaactt 840 ggtattgatt ctatcaagag agtagaaatt ctttctgaag ttcaagctca attgagtgtt 900 gaagctaaag atgtagatgc tcttagtaga actcgtactg ttggtgaagt gattgatgca 960 163 201038734 3240Pro Ala Ser 305 &lt;210&gt; 78 &lt;211&gt; 3303 &lt;212&gt; DNA &lt;213&gt; Thraustochytrium species &lt;400&gt; 78 gcaagtccag ctaccgttcg tgtcgtttca gctcctgttc aagcggctgc tcctgtggg 60 gtatctgctt ctgttgattc tggtttgttg gcaaaagcgg aacaagttgt attggaagta 120 ttggcatcga agactggtta tgagactgag ttgattgaat tggatatgga attggaaact 180 gaacttggta ttgattctat caagagagta gaaattcttt ctgaagttca agctcaattg 240 aatgttgaag ctaaagatgt agatgctctt agtagaactc gtactgttgg tgaagtgatt 300 gatgcaatga aagccgaaat tgctggtggt caaccagctg ctcctgttca agttgcagct 360 cctactcaag tagttgctcc tgttcaagca tctgctcctg ttgattctgg tttgttagca 420 aaagcggaac aagttgtatt ggaagtattg gcatcgaaga ctggttatga gactgagttg 480 attgaattgg atatggaatt ggaaaccgaa cttggtattg attctatcaa gagagtagaa 540 attctttctg aagttcaagc tcaattgagt gttgaagcta Aagatgtaga tgctcttagt 600 agaactcgta ctgttggtga agtgattgat gcaatgaaag ccgaaattgc tggtggtcaa 660 ccagctgctc ctgttcaagt tgcagctcct actcaagtag ttgctcctgt tcaagcatct 720 gctcctgttg attctggttt gttagcaaaa gcggaacaag ttgtatt Gga agtattggca 780 tcgaagactg gttatgagac tgagttgatt gaattggata tggaattgga aaccgaactt 840 ggtattgatt ctatcaagag agtagaaatt ctttctgaag ttcaagctca attgagtgtt 900 gaagctaaag atgtagatgc tcttagtaga actcgtactg ttggtgaagt gattgatgca 960 163 201038734 3240

atgaaagctg aaatttctgg tggtcagcca gctgctcctg ttcaagttgc agctcctact 1020 caaatagttg ctcctgttca agtatccgct cctgttgatt ctggtttgtt agcaaaggcg 1080 gaacaagtag tattggaagt attggcatcc aagactggtt atgagactga gttgattgaa 1140 ttggatatgg aattggaaac tgaacttggt attgattcta tcaagagagt agaaattctt 1200 tctgaagttc aagctcaatt gagtgttgaa gctaaagatg tagatgctct tagtagaact 1260 cgtactgttg gtgaagtgat tgatgcaatg aaagctgaaa tttctggtgg tcaaccaact 1320 gctcctgttc aagttgcagc tcctactcaa atagttgctc ctgttcaagt atctgctcct 1380 gttgattctg gtttgttagc aaaggcggaa caagttgtat tggaagtatt ggcatcgaag 1440 actggttatg agactgagtt gattgaattg gatatggaat tggaaaccga acttggtatt 1500 gattctatca agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct 1560 aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 1620 gccgaaattt ctggtggtca gccagctgct cctgttcaag ttgcagctcc tactcaaata 1680 gttgctcctg ttcaagcatc tgctcctgtt gattctggtt tgttggcaaa agcggaacaa 1740 gttgtattgg aagtgttagc atccaagact ggttatgaaa ctgagttgat tgaattagat 1800 atggaattgg aaaccgaact tggtattgat tctatcaaga gagtagaaat tctttctgaa 1860 gttcaagctc aattgagtgt tgaagctaaa gatgtagatg ctcttagtag aactcgtact 1920 gttggtgaag tgattgatgc aatgaaagct gaaatttctg gtggtcaacc agctgctcct 1980 gttcaagttg cagctcctac tcaaatagtt gctcctgttc aagtatctgc tcctgttgat 2040 tctggtttgt tagcaaaggc ggaacaagtt gtattggaag tattggcatc taagactggt 2100 tatgagactg agttgattga attggatatg gaattggaaa ctgaacttgg tattgattct 2160 atcaagagag tagaaattct ttctgaagtt caagctcaat tgaatgttga agctaaagat 2220 gtagatgctc ttagtagaac tcgtactgtt ggtgaagtga ttgatgcaat gaaagccgaa 2280 attgctggtg gtcaaccagc tgctcctgtt caagttgcag ctcctgctcc agtagttgct 2340 cctgttcaag tatctactcc tgttgattct ggtttgttgg caaaagcgga acaagttgta 2400 ttggaagtgt tagcatgcaa gactggttat gaaactgagt tgattgaatt ggatatggaa 2460 ttggaaactg aacttggtat tgattctatc aagagagtag aaattctttc tgaagttcaa 2520 gctcaattga gtgttgaagc taaagatgta gatgctctta gtagaactcg tactgttggt 2580 gaagtgattg atgcaatgaa agccgaaatt tctggtggtc aaccaactgc tcctgttcaa 2640 gttgcagctc ctactcaagt agttgctcct gttaaagtat ctactcctgt tgattctggt 2700 ttgttagcaa aggcggaaca agtagtattg gaagtattgg catctaagac tggttatgaa 2760 actgagttga ttgaattaga tatggaattg gaaactgaac ttggtattga ttctatcaag 2820 agagtagaaa ttctttctga agttcaagct caattgaatg tggaagctaa agatgtggat 2880 gctcttagta gaactcgtac tgttggtgaa gtgattgatg caatgaaagc cgaaattgct 2940 ggtgatcaac ctgctccagc tgtagttcca gttcaagcta agagtggtgt agccaaccct 3000 gcacttttgg caaaggcgga acaagtagta ttggaagtat tggcatccaa gaccggttat 3060 gaaactgagc tgattgaatt ggatatggaa ttggaaactg aacttggtat tgattcaatc 3120 aagagagtag aaattctgtc cgaagttcaa gcagaattga gtgttgaagc aaaagatgta 3180 gacgctctaa gtagaacccg tactgttggg gaagtgatcg atgcaatgaa agctgaaatt 164 201038734 gctggcagtg ctgtcacggt tgcaactttg gatgattcaa caattatgga ggagacagat gat &lt;210&gt; 79 &lt;211&gt; 1101 &lt;212&gt; PRT &lt;213〉 破囊壺菌物種 &lt;400&gt; 79atgaaagctg aaatttctgg tggtcagcca gctgctcctg ttcaagttgc agctcctact 1020 caaatagttg ctcctgttca agtatccgct cctgttgatt ctggtttgtt agcaaaggcg 1080 gaacaagtag tattggaagt attggcatcc aagactggtt atgagactga gttgattgaa 1140 ttggatatgg aattggaaac tgaacttggt attgattcta tcaagagagt agaaattctt 1200 tctgaagttc aagctcaatt gagtgttgaa gctaaagatg tagatgctct tagtagaact 1260 cgtactgttg gtgaagtgat tgatgcaatg aaagctgaaa tttctggtgg tcaaccaact 1320 gctcctgttc aagttgcagc tcctactcaa atagttgctc ctgttcaagt atctgctcct 1380 gttgattctg gtttgttagc aaaggcggaa caagttgtat tggaagtatt ggcatcgaag 1440 actggttatg agactgagtt gattgaattg gatatggaat tggaaaccga acttggtatt 1500 gattctatca agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct 1560 aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 1620 gccgaaattt ctggtggtca gccagctgct cctgttcaag ttgcagctcc tactcaaata 1680 gttgctcctg ttcaagcatc tgctcctgtt gattctggtt tgttggcaaa agcggaacaa 1740 gttgtattgg aagtgttagc atccaagact ggttatgaaa ctgagttgat tgaattagat 1800 atggaa ttgg aaaccgaact tggtattgat tctatcaaga gagtagaaat tctttctgaa 1860 gttcaagctc aattgagtgt tgaagctaaa gatgtagatg ctcttagtag aactcgtact 1920 gttggtgaag tgattgatgc aatgaaagct gaaatttctg gtggtcaacc agctgctcct 1980 gttcaagttg cagctcctac tcaaatagtt gctcctgttc aagtatctgc tcctgttgat 2040 tctggtttgt tagcaaaggc ggaacaagtt gtattggaag tattggcatc taagactggt 2100 tatgagactg agttgattga attggatatg gaattggaaa ctgaacttgg tattgattct 2160 atcaagagag tagaaattct ttctgaagtt caagctcaat tgaatgttga agctaaagat 2220 gtagatgctc ttagtagaac tcgtactgtt ggtgaagtga ttgatgcaat gaaagccgaa 2280 attgctggtg gtcaaccagc tgctcctgtt caagttgcag ctcctgctcc agtagttgct 2340 cctgttcaag tatctactcc tgttgattct ggtttgttgg caaaagcgga acaagttgta 2400 ttggaagtgt tagcatgcaa gactggttat gaaactgagt tgattgaatt ggatatggaa 2460 ttggaaactg aacttggtat tgattctatc aagagagtag aaattctttc tgaagttcaa 2520 gctcaattga gtgttgaagc taaagatgta gatgctctta gtagaactcg tactgttggt 2580 gaagtgattg atgcaatgaa agccgaaatt tctggtggtc aaccaactgc tcctgttcaa 2640 gttgcagctc c tactcaagt agttgctcct gttaaagtat ctactcctgt tgattctggt 2700 ttgttagcaa aggcggaaca agtagtattg gaagtattgg catctaagac tggttatgaa 2760 actgagttga ttgaattaga tatggaattg gaaactgaac ttggtattga ttctatcaag 2820 agagtagaaa ttctttctga agttcaagct caattgaatg tggaagctaa agatgtggat 2880 gctcttagta gaactcgtac tgttggtgaa gtgattgatg caatgaaagc cgaaattgct 2940 ggtgatcaac ctgctccagc tgtagttcca gttcaagcta agagtggtgt agccaaccct 3000 gcacttttgg caaaggcgga acaagtagta ttggaagtat tggcatccaa gaccggttat 3060 gaaactgagc tgattgaatt ggatatggaa ttggaaactg aacttggtat tgattcaatc 3120 aagagagtag aaattctgtc cgaagttcaa gcagaattga gtgttgaagc aaaagatgta 3180 gacgctctaa gtagaacccg tactgttggg gaagtgatcg atgcaatgaa agctgaaatt 164 201038734 gctggcagtg ctgtcacggt tgcaactttg gatgattcaa caattatgga ggagacagat gat &lt; 210 &gt; 79 &lt; 211 &gt; 1101 &lt; 212 &gt; PRT &lt; 213> Thraustochytrium Species &lt;400&gt; 79

Ala Ser Pro Ala Thr Val Arg Val Val Ser Ala Pro Val Gin Ala Ala 15 10 15Ala Ser Pro Ala Thr Val Arg Val Val Ser Ala Pro Val Gin Ala Ala 15 10 15

Ala Pro Val Gin Val Ser Ala Ser Val Asp Ser Gly Leu Leu Ala Lys 20 25 30Ala Pro Val Gin Val Ser Ala Ser Val Asp Ser Gly Leu Leu Ala Lys 20 25 30

Ala Glu Gin Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr Glu 35 40 45 oAla Glu Gin Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr Glu 35 40 45 o

3300 33033300 3303

Thr Glu Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie 50 55 60Thr Glu Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie 50 55 60

Asp Ser He Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu 65 70 75 80Asp Ser He Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu 65 70 75 80

Asn Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val 85 90 95Asn Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val 85 90 95

Gly Glu Val lie Asp Ala Met Lys Ala Glu lie Ala Gly Gly Gin Pro 100 105 110Gly Glu Val lie Asp Ala Met Lys Ala Glu lie Ala Gly Gly Gin Pro 100 105 110

Ala Ala Pro Val Gin Val Ala Ala Pro Thr Gin Val Val Ala Pro Val 115 120 125Ala Ala Pro Val Gin Val Ala Ala Pro Thr Gin Val Val Ala Pro Val 115 120 125

Gin Ala Ser Ala Pro Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin 130 135 140Gin Ala Ser Ala Pro Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin 130 135 140

Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu 145 150 155 160 lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie 165 170 175Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu 145 150 155 160 lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie 165 170 175

Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu 180 185 190Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu 180 185 190

Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val 195 200 205 工le Asp Ala Met Lys Ala Glu lie Ala Gly Gly Gin Pro Ala Ala Pro 210 215 220Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val 195 200 205 work le Asp Ala Met Lys Ala Glu lie Ala Gly Gly Gin Pro Ala Ala Pro 210 215 220

Val Gin Val Ala Ala Pro Thr Gin Val Val Ala Pro Val Gin Ala Ser 225 230 235 240Val Gin Val Ala Ala Pro Thr Gin Val Val Ala Pro Val Gin Ala Ser 225 230 235 240

Ala Pro Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu 245 250 255 165 201038734 u e L u _—t G e o 1 7 12 u e L u T G r h T u &lt;—I G r 5 y6 T 2Ala Pro Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu 245 250 255 165 201038734 u e L u _-t G e o 1 7 12 u e L u T G r h T u &lt;-I G r 5 y6 T 2

Glu Val Leu Ala Ser Lys Thr Gly 260Glu Val Leu Ala Ser Lys Thr Gly 260

Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val 275 280 285Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val 275 280 285

Glu lie Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp 290 295 300Glu lie Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp 290 295 300

Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala 305 310 315 320Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala 305 310 315 320

Met Lys Ala Glu lie Ser Gly Gly Gin Pro Ala Ala Pro Val Gin Val 325 330 335Met Lys Ala Glu lie Ser Gly Gly Gin Pro Ala Ala Pro Val Gin Val 325 330 335

r o h 4 T 3 〇 r p a I—I A a I—_ Ar o h 4 T 3 〇 r p a I—I A a I—_ A

Gin lie Val Ala Pro Val Gin Val Ser Ala Pro Val 345 350Gin lie Val Ala Pro Val Gin Val Ser Ala Pro Val 345 350

Asp Ser Gly Leu 355Asp Ser Gly Leu 355

Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val Leu 360 365 oLeu Ala Lys Ala Glu Gin Val Val Leu Glu Val Leu 360 365 o

s Ly r o e 7 s 3 a I—_ As Ly r o e 7 s 3 a I—_ A

Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met Glu 375 380Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met Glu 375 380

Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu 385 390 395 400Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu 385 390 395 400

Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp Ala 405 410 415Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp Ala 405 410 415

Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys Ala 420 425 430Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys Ala 420 425 430

Glu lie Ser Gly Gly Gin Pro Thr Ala Pro Val Gin Val Ala Ala Pro 435 440 445Glu lie Ser Gly Gly Gin Pro Thr Ala Pro Val Gin Val Ala Ala Pro 435 440 445

no 11.0 G 4 r h TNo 11.0 G 4 r h T

e _—_ Ie _—_ I

a Va V

a V n I—_ G I—I L〇 a 5 V4 o r p a -—_ Aa V n I—_ G I—I L〇 a 5 V4 o r p a ——_ A

Ser Ala Pro Val Asp Ser Gly 460Ser Ala Pro Val Asp Ser Gly 460

Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val Leu Ala Ser Lys 465 470 475 480Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val Leu Ala Ser Lys 465 470 475 480

Thr Gly Tyr Glu Thr Glu Leu He Glu Leu Asp Met Glu Leu Glu Thr 485 490 495Thr Gly Tyr Glu Thr Glu Leu He Glu Leu Asp Met Glu Leu Glu Thr 485 490 495

Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val 500 505 510Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val 500 505 510

Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg 515 520 525Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg 515 520 525

Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys Ala Glu lie Ser 530 535 540Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys Ala Glu lie Ser 530 535 540

Gly Gly Gin Pro Ala Ala Pro Val Gin Val Ala Ala Pro Thr Gin He 166 201038734 545 550 555 560Gly Gly Gin Pro Ala Ala Pro Val Gin Val Ala Ala Pro Thr Gin He 166 201038734 545 550 555 560

Val Ala Pro Val Gin Ala Ser Ala Pro Val Asp Ser Gly Leu Leu Ala 565 570 575Val Ala Pro Val Gin Ala Ser Ala Pro Val Asp Ser Gly Leu Leu Ala 565 570 575

Lys Ala Glu Gin Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr 580 585 590Lys Ala Glu Gin Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr 580 585 590

Glu Thr Glu Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly 595 600 605 工le Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Gin 610 615 620Glu Thr Glu Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly 595 600 605 work le Asp Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Gin 610 615 620

Leu Ser Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr 625 630 635 640 ❹Leu Ser Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr 625 630 635 640 ❹

Val Gly Glu Val 工le Asp Ala Met Lys Ala Glu lie Ser Gly Gly Gin 645 650 655Val Gly Glu Val work le Asp Ala Met Lys Ala Glu lie Ser Gly Gly Gin 645 650 655

Pro Ala Ala Pro Val Gin Val Ala Ala Pro Thr Gin lie Val Ala Pro 660 665 670Pro Ala Ala Pro Val Gin Val Ala Ala Pro Thr Gin lie Val Ala Pro 660 665 670

Val Gin Val Ser Ala Pro Val Asp Ser Gly Leu Leu Ala Lys Ala Glu 67 5 680 685Val Gin Val Ser Ala Pro Val Asp Ser Gly Leu Leu Ala Lys Ala Glu 67 5 680 685

Gin Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu 690 695 700Gin Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu 690 695 700

Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser 705 710 715 720 lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu Asn Val 725 730 735Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser 705 710 715 720 lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu Asn Val 725 730 735

Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu 740 745 750Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu 740 745 750

Val lie Asp Ala Met Lys Ala Glu lie Ala Gly Gly Gin Pro Ala Ala 755 760 765Val lie Asp Ala Met Lys Ala Glu lie Ala Gly Gly Gin Pro Ala Ala 755 760 765

Pro Val Gin Val Ala Ala Pro Ala Pro Val Val Ala Pro Val Gin Val 770 775 780Pro Val Gin Val Ala Ala Pro Ala Pro Val Val Ala Pro Val Gin Val 770 775 780

Ser Thr Pro Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val 785 790 795 800Ser Thr Pro Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val 785 790 795 800

Leu Glu Val Leu Ala Cys Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu 805 810 815Leu Glu Val Leu Ala Cys Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu 805 810 815

Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg 820 . 825 830Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg 820 . 825 830

Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys 835 840 845 167 201038734Val Glu lie Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys 835 840 845 167 201038734

Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp 850 855 860Asp Val Asp Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp 850 855 860

Ala Met Lys Ala Glu lie Ser Gly Gly Gin 865 870 a V o r p a t—I A r h T o 5 r 7 p 8 η o 18 G 8Ala Met Lys Ala Glu lie Ser Gly Gly Gin 865 870 a V o r p a t—I A r h T o 5 r 7 p 8 η o 18 G 8

a Va V

A _—I a V _—t a V n _—_ G r 5 h 8 T 8 〇 r p a _—_ A a I~~_ A a V s y L.—_ a V 〇o r 9 p 8 〇 r p r 5 h 9 T 8 r e sA _—I a V _—ta V n _—_ G r 5 h 8 T 8 〇rpa _—_ A a I~~_ A a V sy L.—_ a V 〇or 9 p 8 〇rpr 5 h 9 T 8 res

Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 900 905 910Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 900 905 910

Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 915 920 925Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 915 920 925

Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 930 935 940 aGlu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 930 935 940 a

Leu Ser Glu Val Gin Ala Gin Leu Asn Val Glu Ala Lys Asp Val Asp 945 950 955 960Leu Ser Glu Val Gin Ala Gin Leu Asn Val Glu Ala Lys Asp Val Asp 945 950 955 960

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val 工le Asp Ala Met Lys 965 970 975Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val work Le Asp Ala Met Lys 965 970 975

Ala Glu lie Ala Gly Asp Gin Pro Ala Pro Ala Val Val Pro Val Gin 980 985 990Ala Glu lie Ala Gly Asp Gin Pro Ala Pro Ala Val Val Pro Val Gin 980 985 990

Ala Lys Ser Gly Val Ala Asn Pro Ala Leu Leu Ala Lys Ala Glu Gin 995 1000 1005Ala Lys Ser Gly Val Ala Asn Pro Ala Leu Leu Ala Lys Ala Glu Gin 995 1000 1005

Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu 1010 1015 1020Val Val Leu Glu Val Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu 1010 1015 1020

Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp 1025 1030 1035Leu lie Glu Leu Asp Met Glu Leu Glu Thr Glu Leu Gly lie Asp 1025 1030 1035

Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Glu Leu 1040 1045 1050 1.)Ser lie Lys Arg Val Glu lie Leu Ser Glu Val Gin Ala Glu Leu 1040 1045 1050 1.)

Ser Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr 1055 1060 1065Ser Val Glu Ala Lys Asp Val Asp Ala Leu Ser Arg Thr Arg Thr 1055 1060 1065

Val Gly Glu Val lie Asp Ala Met Lys Ala Glu lie Ala Gly Ser 1070 1075 1080Val Gly Glu Val lie Asp Ala Met Lys Ala Glu lie Ala Gly Ser 1070 1075 1080

Ala Val Thr Val Ala Thr Leu Asp Asp Ser Thr lie Met Glu Glu 1085 1090 1095Ala Val Thr Val Ala Thr Leu Asp Asp Ser Thr lie Met Glu Glu 1085 1090 1095

Thr Asp Asp 1100 &lt;210&gt; 80 &lt;211〉 255 &lt;212&gt; DNA &lt;213〉 破囊壺菌物種 &lt;400&gt; 80 gttgattctg gtttgttggc aaaagcggaa caagttgtat tggaagtatt ggcatcgaag 168 60 201038734 actggttatg gattctatca aaagatgtag gccgaaattg agactgagtt agagagtaga atgctcttag ctggt gattgaattg aattctttct tagaactcgt gatatggaat gaagttcaag actgttggtg tggaaactga ctcaattgaa aagtgattga acttggtatt tgttgaagct tgcaatgaaa 120 180 240 255 &lt;210〉 81 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213〉 破囊壺菌物種 &lt;400&gt; 81Thr Asp Asp 1100 &lt;210&gt; 80 &lt;211> 255 &lt;212&gt; DNA &lt;213> Thraustochytrium Species &lt;400&gt; 80 gttgattctg gtttgttggc aaaagcggaa caagttgtat tggaagtatt ggcatcgaag 168 60 201038734 actggttatg gattctatca aaagatgtag gccgaaattg agactgagtt agagagtaga atgctcttag ctggt Gattgaattg aattctttct tagaactcgt gatatggaat gaagttcaag actgttggtg tggaaactga ctcaattgaa aagtgattga acttggtatt tgttgaagct tgcaatgaaa 120 180 240 255 &lt;210> 81 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213> Thraustochytrium species &lt;400&gt;

Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15

Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30

Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45

Leu Ser Glu Val Gin Ala Gin Leu Asn Val Glu Ala Lys Asp Val Asp 50 55 60Leu Ser Glu Val Gin Ala Gin Leu Asn Val Glu Ala Lys Asp Val Asp 50 55 60

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80

Ala Glu He Ala Gly 85 &lt;210〉 82 &lt;211&gt; 255 &lt;212&gt; DNA &lt;213〉 破囊壺菌物種 &lt;400&gt; 82 60 120 180 240 255 gttgattctg gtttgttagc aaaagcggaa caagttgtat tggaagtatt ggcatcgaag actggttatg agactgagtt gattgaattg gatatggaat tggaaaccga acttggtatt gattctatca agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa gccgaaattg ctggt &lt;210&gt; 83 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 83Ala Glu He Ala Gly 85 &lt;210> 82 &lt;211&gt; 255 &lt;212&gt; DNA &lt;213> Thraustochytrium species &lt;400&gt; 82 60 120 180 240 255 gttgattctg gtttgttagc aaaagcggaa caagttgtat tggaagtatt ggcatcgaag actggttatg agactgagtt gattgaattg gatatggaat Tggaaaccga acttggtatt gattctatca agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa gccgaaattg ctggt &lt;210&gt; 83 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213&gt; Thraustochytrium species &lt;400&gt; 83

Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15

Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30

Glu Leu Glu Thr Glu Leu Gly 工le Asp Ser lie Lys Arg Val Glu lie 35 40 45 169 201038734Glu Leu Glu Thr Glu Leu Gly work le Asp Ser lie Lys Arg Val Glu lie 35 40 45 169 201038734

Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val 工le Asp Ala Met Lys 65 70 75 80Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val work le Asp Ala Met Lys 65 70 75 80

Ala Glu lie Ala Gly 85 &lt;210&gt; 84 &lt;211&gt; 252 &lt;212&gt; DNA &lt;213〉 破囊壺g物種 &lt;400&gt; 84 gttgattctg gtttgttagc aaaagcggaa caagttgtat tggaagtatt ggcatcgaag 60 actggttatg agactgagtt gattgaattg gatatggaat tggaaaccga acttggtatt 120Ala Glu lie Ala Gly 85 &lt;210&gt; 84 &lt;211&gt; 252 &lt;212&gt; DNA &lt;213> Thrausally pot g species &lt;400&gt; 84 gttgattctg gtttgttagc aaaagcggaa caagttgtat tggaagtatt ggcatcgaag 60 actggttatg agactgagtt gattgaattg gatatggaat tggaaaccga acttggtatt 120

gattctatca agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct 180 aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 240 gctgaaattt ct 252 &lt;210〉 85 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;400〉 85Gattctatca agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct 180 aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 240 gctgaaattt ct 252 &lt;210> 85 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213&gt; Thraustochytrium species &lt;400> 85

Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15

Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30

Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45

Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80

Ala Glu lie Ser Gly 85 &lt;210〉 86 &lt;211&gt; 255 &lt;212&gt; DNA &lt;213〉 破囊壺菌物種 &lt;400&gt; 86 gttgattctg gtttgttagc aaaggcggaa caagtagtat tggaagtatt ggcatccaag 60 actggttatg agactgagtt gattgaattg gatatggaat tggaaactga acttggtatt 120 gattctatca agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct 180 aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 240 170 201038734 255 gctgaaattt ctggt &lt;210&gt; 87 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 87Ala Glu lie Ser Gly 85 &lt;210> 86 &lt;211&gt; 255 &lt;212&gt; DNA &lt;213> Thraustochytrium species &lt;400&gt; 86 gttgattctg gtttgttagc aaaggcggaa caagtagtat tggaagtatt ggcatccaag 60 actggttatg agactgagtt gattgaattg gatatggaat tggaaactga acttggtatt 120 gattctatca Agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct 180 aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 240 170 201038734 255 gctgaaattt ctggt &lt;210&gt; 87 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213&gt; Thraustochytrium species &lt;400&gt;

Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15

Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30

Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45

Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80

Ala Glu lie Ser Gly 85 . &lt;210&gt; 88 一 &lt;211&gt; 254Ala Glu lie Ser Gly 85 . &lt;210&gt; 88 one &lt;211&gt; 254

- &lt;212&gt; DNA &lt;213&gt; 破囊壺菌物種 * &lt;400&gt; 88 60 120 180 240 254 ttgattctgg tttgttagca aaggcggaac aagttgtatt ggaagtattg gcatcgaaga ctggttatga gactgagttg attgaattgg atatggaatt ggaaaccgaa cttggtattg attctatcaa gagagtagaa attctttctg aagttcaagc tcaattgagt gttgaagcta aagatgtaga tgctcttagt agaactcgta ctgttggtga agtgattgat gcaatgaaag ccgaaatttc tggt 〇 &lt;210&gt; 89 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 89- &lt; 212 &gt; DNA &lt; 213 &gt; Thraustochytrium species * &lt; agtgattgat gcaatgaaag 88 60 120 180 240 254 ttgattctgg tttgttagca aaggcggaac aagttgtatt ggaagtattg gcatcgaaga ctggttatga gactgagttg attgaattgg atatggaatt ggaaaccgaa cttggtattg attctatcaa gagagtagaa attctttctg aagttcaagc tcaattgagt gttgaagcta aagatgtaga tgctcttagt agaactcgta ctgttggtga; 400 & gt Ccgaaatttc tggt 〇&lt;210&gt; 89 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213&gt; Thraustochytrium species &lt;400&gt; 89

Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15

Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30

Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45

Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60 171 201038734Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60 171 201038734

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val 工le Asp Ala Met Lys 65 70 75 80Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val work le Asp Ala Met Lys 65 70 75 80

Ala Glu lie Ser Gly 85 &lt;210&gt; 90 &lt;211〉 255 &lt;212&gt; DNA &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 90 gttgattctg gtttgttggc aaaagcggaa caagttgtat tggaagtgtt agcatccaag 60 actggttatg aaactgagtt gattgaatta gatatggaat tggaaaccga acttggtatt 120 gattctatca agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct 180 aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 240 gctgaaattt ctggt 255Ala Glu lie Ser Gly 85 &lt;210&gt; 90 &lt;211&gt; 255 &lt;212&gt; DNA &lt;213&gt; Thraustochytrium species &lt;400&gt; 90 gttgattctg gtttgttggc aaaagcggaa caagttgtat tggaagtgtt agcatccaag 60 actggttatg aaactgagtt gattgaatta gatatggaat tggaaaccga acttggtatt 120 gattctatca Agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct 180 aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 240 gctgaaattt ctggt 255

&lt;21〇&gt; 91 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 91&lt;21〇&gt; 91 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213&gt; Thraustochytrium species &lt;400&gt;

Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15

Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30

Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45

Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80

Ala Glu lie Ser Gly 85 &lt;210&gt; 92 &lt;211〉 255 &lt;212&gt; DNA &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 92 gttgattctg gtttgttagc aaaggcggaa caagttgtat tggaagtatt ggcatctaag 60 actggttatg agactgagtt gattgaattg gatatggaat tggaaactga acttggtatt 120 gattctatca agagagtaga aattctttct gaagttcaag ctcaattgaa tgttgaagct 180 aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 240 gccgaaattg ctggt 255 &lt;210&gt; 93 &lt;211&gt; 85 172 201038734 &lt;212&gt; PRT &lt;213〉 破囊壺菌物種 &lt;400&gt; 93Ala Glu lie Ser Gly 85 &lt;210&gt; 92 &lt;211&gt; 255 &lt;212&gt; DNA &lt;213&gt; Thraustochytrium species &lt;400&gt; 92 gttgattctg gtttgttagc aaaggcggaa caagttgtat tggaagtatt ggcatctaag 60 actggttatg agactgagtt gattgaattg gatatggaat tggaaactga acttggtatt 120 gattctatca Agagagtaga aattctttct gaagttcaag ctcaattgaa tgttgaagct 180 aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 240 gccgaaattg ctggt 255 &lt;210&gt; 93 &lt;211&gt; 85 172 201038734 &lt;212&gt; PRT &lt;213> Thraustochytrium species &lt;400&gt;

Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15

Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30

Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45

Leu Ser Glu Val Gin Ala Gin Leu Asn Val Glu Ala Lys Asp Val Asp 50 55 60Leu Ser Glu Val Gin Ala Gin Leu Asn Val Glu Ala Lys Asp Val Asp 50 55 60

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80

Ala Glu lie Ala Gly 85 &lt;210&gt; 94 &lt;211&gt; 255 &lt;212&gt; DNA &lt;213〉 破囊壺菌物種 &lt;400&gt; 94 60 120 180 240 255 gttgattctg gtttgttggc aaaagcggaa caagttgtat tggaagtgtt agcatgcaag actggttatg aaactgagtt gattgaattg gatatggaat tggaaactga acttggtatt gattctatca agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa gccgaaattt ctggt &lt;210&gt; 95 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213〉破囊壺菌物種 &lt;400&gt; 95Ala Glu lie Ala Gly 85 &lt;210&gt; 94 &lt;211&gt; 255 &lt;212&gt; DNA &lt;213&gt; Thraustochytrium species &lt;400&gt; 94 60 120 180 240 255 gttgattctg gtttgttggc aaaagcggaa caagttgtat tggaagtgtt agcatgcaag actggttatg aaactgagtt gattgaattg gatatggaat Tggaaactga acttggtatt gattctatca agagagtaga aattctttct gaagttcaag ctcaattgag tgttgaagct aaagatgtag atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa gccgaaattt ctggt &lt;210&gt; 95 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213> Thraustochytrium species &lt;400&gt;

Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15

Leu Ala Cys Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30Leu Ala Cys Lys Thr Gly Tyr Glu Thr Glu Leu lie Glu Leu Asp Met 20 25 30

Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45

Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60Leu Ser Glu Val Gin Ala Gin Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80

Ala Glu lie Ser Gly 85 173 201038734 &lt;210&gt; 96 &lt;211〉 255 &lt;212&gt; DNA &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 96 gttgattctg gtttgttagc aaaggcggaa caagtagtat tggaagtatt ggcatctaag 60 actggttatg aaactgagtt gattgaatta gatatggaat tggaaactga acttggtatt 120 gattctatca agagagtaga aattctttct gaagttcaag ctcaattgaa tgtggaagct 180 aaagatgtgg atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 240 gccgaaattg ctggt 255 &lt;210&gt; 97 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 97Ala Glu lie Ser Gly 85 173 201038734 &lt;210&gt; 96 &lt;211&gt; 255 &lt;212&gt; DNA &lt;213&gt; Thraustochytrium species &lt;400&gt; 96 gttgattctg gtttgttagc aaaggcggaa caagtagtat tggaagtatt ggcatctaag 60 actggttatg aaactgagtt gattgaatta gatatggaat tggaaactga acttggtatt 120 gattctatca agagagtaga aattctttct gaagttcaag ctcaattgaa tgtggaagct 180 aaagatgtgg atgctcttag tagaactcgt actgttggtg aagtgattga tgcaatgaaa 240 gccgaaattg ctggt 255 &lt;210&gt; 97 &lt;211&gt; 85 &lt;212&gt; PRT &lt;213&gt; Thraustochytrium species &lt;400&gt;

Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15Val Asp Ser Gly Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 15 10 15

Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu 工le Glu Leu Asp Met 20 25 30Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu work le Glu Leu Asp Met 20 25 30

Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie Lys Arg Val Glu lie 35 40 45

Leu Ser Glu Val Gin Ala Gin Leu Asn Val Glu Ala Lys Asp Val Asp 50 55 60Leu Ser Glu Val Gin Ala Gin Leu Asn Val Glu Ala Lys Asp Val Asp 50 55 60

Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80Ala Leu Ser Arg Thr Arg Thr Val Gly Glu Val lie Asp Ala Met Lys 65 70 75 80

Ala Glu He Ala Gly 85 &lt;210&gt; 98 &lt;211&gt; 255 &lt;212&gt; DNA &lt;213〉 破袭壺菌物種 &lt;400&gt; 98 gccaaccctg cacttttggc aaaggcggaa caagtagtat tggaagtatt ggcatccaag 60 accggttatg aaactgagct gattgaattg gatatggaat tggaaactga acttggtatt 120 gattcaatca agagagtaga aattctgtcc gaagttcaag cagaattgag tgttgaagca 180 aaagatgtag acgctctaag tagaacccgt actgttgggg aagtgatcga tgcaatgaaa 240 gctgaaattg ctgqc 255 &lt;210〉 &lt;211&gt; 種 物 菌 壺 o &lt;212&gt; &lt;213〉 &lt;400&gt; 99Ala Glu He Ala Gly 85 &lt;210&gt; 98 &lt;211&gt; 255 &lt;212&gt; DNA &lt;213> Spoiled chytrid species &lt;400&gt; 98 gccaaccctg cacttttggc aaaggcggaa caagtagtat tggaagtatt ggcatccaag 60 accggttatg aaactgagct gattgaattg gatatggaat tggaaactga acttggtatt 120 gattcaatca Agagagtaga aattctgtcc gaagttcaag cagaattgag tgttgaagca 180 aaagatgtag acgctctaag tagaacccgt actgttgggg aagtgatcga tgcaatgaaa 240 gctgaaattg ctgqc 255 &lt;210> &lt;211&gt; species bacteria pot o &lt;212&gt;&lt;213&gt;&lt;400&gt;

Ala Asn Pro Ala Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 174 201038734 5 10 15 lie Glu Leu Asp Met 30Ala Asn Pro Ala Leu Leu Ala Lys Ala Glu Gin Val Val Leu Glu Val 174 201038734 5 10 15 lie Glu Leu Asp Met 30

Lys Arg Val Glu lie 45Lys Arg Val Glu lie 45

Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu 20 25Leu Ala Ser Lys Thr Gly Tyr Glu Thr Glu Leu 20 25

Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie 35 40Glu Leu Glu Thr Glu Leu Gly lie Asp Ser lie 35 40

Leu Ser Glu Val Gin Ala Glu Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60Leu Ser Glu Val Gin Ala Glu Leu Ser Val Glu Ala Lys Asp Val Asp 50 55 60

Ala Leu Ser Arg Thr Ar 65 70 Ala Glu lie Ala Gly 85 &lt;210&gt; 100 &lt;211&gt; 2151 &lt;212&gt; DNA &lt;213〉 破囊壺菌物種 &lt;4〇〇&gt; 100 gtttcaagca ttgaatcttc gatagcgact atggtatgca tccctcaacg acccgattaa ggtaaacttg gtatggacaa atcgccgaac gtggtgctat gtttttgctc gtggtgttga attgttgatg agatttcttg ggagaaagat tcacaactga aagaagaagg atgctttcct cgtgaaattg ctaaagcagt gctgatgaac ccttgtgggc gcatttttga aggaagagtt aaatctttga ttgataaagt gaagcccatg gagcaaaagc aaggctgcag tgcaaaaagt gcatcaggcg ttttgaggga gtatatggaa ccaaagtaac gttcgtcatt tggttatgtt gattatgcaa tggctaacga tctcaattgt gtgttaaatc gctttgaaaa aacaatttca gagactgttg caagaatagt ggtgttcctc cagtttcacc70 Ala Glu lie Ala Gly 85 &lt;210&gt; 100 &lt tccctcaacg acccgattaa ggtaaacttg gtatggacaa atcgccgaac gtggtgctat gtttttgctc gtggtgttga attgttgatg agatttcttg ggagaaagat tcacaactga aagaagaagg atgctttcct cgtgaaattg ctaaagcagt gctgatgaac ccttgtgggc gcatttttga aggaagagtt aaatctttga ttgataaagt gaagcccatg gagcaaaagc aaggctgcag tgcaaaaagt gcatcaggcg ttttgaggga gtatatggaa ccaaagtaac gttcgtcatt tggttatgtt gattatgcaa tggctaacga tctcaattgt gtgttaaatc gctttgaaaa aacaatttca gagactgttg caagaatagt ggtgttcctc cagtttcacc

Thr Val Gly Glu Val 75 gtatggaaaa attggtggct acttggatgg gcgttaatgg gaatggaaga accttctttt tgcttcagtt catgatcaag ctttggtttg tgcaaaactt tattgctgaa ggtatgagtt tgcaaatctt tccattcggg agctcacaaa ttggttactg agtatctggt ggtgctcgtg gaaaggtggc acttacattt taatggtaaa tccggaaaag tgcagctggg cgtggtagta gctcggtatt agggaggtta tatatatttg tcttgcgatg tgaaaaggag catctagttc taaattggtt gagaacaaaa tggactagta aacttgctgt tagttctttg gctggatatc atcacttaac aagattggtt tatttgtttt ggaccttggg atcaatgggt gtccagatta cttatcttca aatccttctc tttgtcaaaa tcggcaacta 工le Asp Ala Met Lys 80 ttgtttatca acattttcat cagcgaaaca tttgaaagag tggctgttgc gcgtatgaat gaatagtgga atcatgcggt tggatttgga atggcctaat atagtttggc tgcggaattg aatctggtta cacgattagc gaaagcctca tgctccgatt gtattactcc actttgtatt tgatgggtcg atcagctttg atttagataa agctggtttg aaccaactcc aaaagttcac gagcatctat tgcaaatata tatcttccgc tgagaaagta gtattactgg tattgtgcat ctttggatga tttcaacgca cagcagtgaa catgaatttt atggaaatgt tggtcaatct ttagattggg tgcagcttat atggtggaat ggtaactcca ttcctcgtga aggtggcgcg aagttttagt tggcaactgg ttgttcaaac ttttacccct 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 175 1380 201038734 gagttaaatc catttctaaa gtctcatcaa attcatggta aaaatgtttt gcctatgact 1440 gtagcaattg gatatcttgc tcacttggtt aagaattttt atgctggtca tcatttgtgg 1500 ggagttgaag atgctcaatt gttcagtggt gttgtaattg accatgcggt gcaagctcaa 1560 gtgaaattaa cggaacagag tttggatgat gatggcaagg taaaagttca agctgttctg 1620 actgcttcaa acgataatgg aaaaatggta cctgcataca aagcagtgat tgttttggga 1680 aaaacaagta gacctgcgtt tattttgaaa gatttttcat tgcaagaatc taattctcgc 1740 agtgctgatg agttgtatga tggtaaaact ttgtttcatg gtccattatt tcgtggaatt 1800 accaagttgt tgaatgtatc tgatacttca ctaacaactc aatgtaccaa tattgatttg 1860 actgctactg aacgtggtca atttgcggat atcgaacctg tgaatccttt tatggcggat 1920 gctgcatttc aagctatgct tgtatgggtt agaaatttaa ggaatagtgc atctttacca 1980 aacaattgtg aaagagtaga tatctataaa ccaatagcac ctggtgaaaa gtattacact 2040 actttgcaag ctttgggtaa tacctccggt tctgttctca agtctgtatt ttatatgcac 2100Thr Val Gly Glu Val 75 gtatggaaaa attggtggct acttggatgg gcgttaatgg gaatggaaga accttctttt tgcttcagtt catgatcaag ctttggtttg tgcaaaactt tattgctgaa ggtatgagtt tgcaaatctt tccattcggg agctcacaaa ttggttactg agtatctggt ggtgctcgtg gaaaggtggc acttacattt taatggtaaa tccggaaaag tgcagctggg cgtggtagta gctcggtatt agggaggtta tatatatttg tcttgcgatg tgaaaaggag catctagttc taaattggtt gagaacaaaa tggactagta aacttgctgt tagttctttg gctggatatc atcacttaac aagattggtt tatttgtttt ggaccttggg atcaatgggt gtccagatta cttatcttca aatccttctc tttgtcaaaa tcggcaacta work le Asp Ala Met Lys 80 ttgtttatca acattttcat cagcgaaaca tttgaaagag tggctgttgc gcgtatgaat gaatagtgga atcatgcggt tggatttgga atggcctaat atagtttggc tgcggaattg aatctggtta cacgattagc gaaagcctca tgctccgatt gtattactcc actttgtatt tgatgggtcg atcagctttg atttagataa agctggtttg aaccaactcc aaaagttcac gagcatctat tgcaaatata tatcttccgc tgagaaagta gtattactgg tattgtgcat ctttggatga tttcaacgca cagcagtgaa catgaatttt atggaaatgt tggtcaatct ttagattggg tgcagcttat atggtggaat ggtaactcca ttcctc gtga aggtggcgcg aagttttagt tggcaactgg ttgttcaaac ttttacccct 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 175 1380 201038734 gagttaaatc catttctaaa gtctcatcaa attcatggta aaaatgtttt gcctatgact 1440 gtagcaattg gatatcttgc tcacttggtt aagaattttt atgctggtca tcatttgtgg 1500 ggagttgaag atgctcaatt gttcagtggt gttgtaattg accatgcggt gcaagctcaa 1560 gtgaaattaa cggaacagag tttggatgat gatggcaagg taaaagttca agctgttctg 1620 actgcttcaa acgataatgg aaaaatggta cctgcataca aagcagtgat tgttttggga 1680 aaaacaagta gacctgcgtt tattttgaaa gatttttcat tgcaagaatc taattctcgc 1740 agtgctgatg agttgtatga tggtaaaact ttgtttcatg gtccattatt tcgtggaatt 1800 accaagttgt tgaatgtatc tgatacttca ctaacaactc aatgtaccaa tattgatttg 1860 actgctactg aacgtggtca atttgcggat atcgaacctg tgaatccttt tatggcggat 1920 gctgcatttc aagctatgct tgtatgggtt agaaatttaa ggaatagtgc atctttacca 1980 aacaattgtg aaagagtaga tatctataaa ccaatagcac ctggtgaaaa gtattacact 2040 actttgcaag ctttgggtaa tacctccggt tctgttctca agt Ctgtatt ttatatgcac 2100

gatgaacaag gagaagtatt tctatctgga agagctagtg ttgttgtgaa t 2151 &lt;210&gt; 101 &lt;211&gt; 717 &lt;212&gt; PRT &lt;213〉 破囊壺菌物種 &lt;400&gt; 101Gatgaacaag gagaagtatt tctatctgga agagctagtg ttgttgtgaa t 2151 &lt;210&gt; 101 &lt;211&gt; 717 &lt;212&gt; PRT &lt;213> Thraustochytrium species &lt;400&gt; 101

Val Ser Ser lie Glu Ser Ser Tyr Gly Lys lie Gly Gly Phe Val Tyr 15 10 15Val Ser Ser lie Glu Ser Ser Tyr Gly Lys lie Gly Gly Phe Val Tyr 15 10 15

Gin His Phe His Asp Ser Asp Tyr Gly Met Gin Leu Gly Trp Ala Leu 20 25 30Gin His Phe His Asp Ser Asp Tyr Gly Met Gin Leu Gly Trp Ala Leu 20 25 30

Met Ala Ala Lys His Leu Lys Glu Ser Leu Asn Asp Pro lie Lys Asn 35 40 45Met Ala Ala Lys His Leu Lys Glu Ser Leu Asn Asp Pro lie Lys Asn 35 40 45

Gly Arg Thr Phe Phe Leu Ala Val Ala Arg Met Asn Gly Lys Leu Gly 50 55 60Gly Arg Thr Phe Phe Leu Ala Val Ala Arg Met Asn Gly Lys Leu Gly 50 55 60

Met Asp Asn Ala Ser Val His Asp Gin Gly lie Val Glu Ser Cys Gly 65 70 75 80 lie Ala Glu Arg Gly Ala lie Phe Gly Leu Cys Lys Thr Leu Asp Leu 85 90 95Met Asp Asn Ala Ser Val His Asp Gin Gly lie Val Glu Ser Cys Gly 65 70 75 80 lie Ala Glu Arg Gly Ala lie Phe Gly Leu Cys Lys Thr Leu Asp Leu 85 90 95

Glu Trp Pro Asn Val Phe Ala Arg Gly Val Asp lie Ala Glu Gly Met 100 105 110Glu Trp Pro Asn Val Phe Ala Arg Gly Val Asp lie Ala Glu Gly Met 100 105 110

Ser Tyr Ser Leu Ala Ala Glu Leu 工le Val Asp Glu lie Ser Cys Ala 115 120 125Ser Tyr Ser Leu Ala Ala Glu Leu work Le Val Asp Glu lie Ser Cys Ala 115 120 125

Asn Leu Ser lie Arg Glu Ser Gly Tyr Thr lie Ser Gly Glu Arg Phe 130 135 140Asn Leu Ser lie Arg Glu Ser Gly Tyr Thr lie Ser Gly Glu Arg Phe 130 135 140

Thr Thr Glu Ala His Lys Leu Val Thr Gly Lys Pro His Ala Pro lie 145 150 155 160Thr Thr Glu Ala His Lys Leu Val Thr Gly Lys Pro His Ala Pro lie 150 15 155 160

Lys Lys Lys Asp Ala Phe Leu Val Ser Gly Gly Ala Arg Gly lie Thr 176 201038734 165 170 175Lys Lys Lys Asp Ala Phe Leu Val Ser Gly Gly Ala Arg Gly lie Thr 176 201038734 165 170 175

Pro Leu Cys He Arg Glu He Ala Lys Ala Val Lys Gly Gly Thr Tyr 180 185 190 lie Leu Met Gly Arg Ser Ala Leu Ala Asp Glu Pro Leu Trp Ala Asn 195 200 205Pro Leu Cys He Arg Glu He Ala Lys Ala Val Lys Gly Gly Thr Tyr 180 185 190 lie Leu Met Gly Arg Ser Ala Leu Ala Asp Glu Pro Leu Trp Ala Asn 195 200 205

Gly Lys Ser Gly Lys Asp Leu Asp Lys Ala Gly Leu Ala Phe Leu Lys 210 215 220Gly Lys Ser Gly Lys Asp Leu Asp Lys Ala Gly Leu Ala Phe Leu Lys 210 215 220

Glu Glu Phe A!a Ala Gly Arg Gly Ser Lys Pro Thr Pro Lys Val HisGlu Glu Phe A!a Ala Gly Arg Gly Ser Lys Pro Thr Pro Lys Val His

Lys Ser Leu lie Asp Lys Val Leu Gly lie Arg Glu Val Arg Ala Ser 245 250 255 ❹ lie Ala Asn lie Glu Ala His Gly Ala Lys Ala lie Tyr Leu Ser Cys 260 265 270s Ala Asn lie Glu Ala His Gly Ala Lys Ala lie Tyr Leu Ser Cys 260 265 270

Asp Val Ser Ser Ala Glu Lys Val Lys Ala Ala Val Gin Lys Val Glu 275 280 285Asp Val Ser Ser Ala Glu Lys Val Lys Ala Ala Val Gin Lys Val Glu 275 280 285

Lys Glu His Leu Val Arg lie Thr Gly lie Val His Ala Ser Gly Val 290 295 300Lys Glu His Leu Val Arg lie Thr Gly lie Val His Ala Ser Gly Val 290 295 300

Leu Arg Asp Lys Leu Val Glu Asn Lys Thr Leu Asp Asp Phe Asn Ala 305 310 315 320Leu Arg Asp Lys Leu Val Glu Asn Lys Thr Leu Asp Asp Phe Asn Ala 305 310 315 320

Val Tyr Gly Thr Lys Val Thr Gly Leu Val Asn Leu Leu Ser Ala Val 325 330 335Val Tyr Gly Thr Lys Val Thr Gly Leu Val Asn Leu Leu Ser Ala Val 325 330 335

Asn Met Asn Phe Val Arg His Leu Val Met Phe Ser Ser Leu Ala Gly 340 345 350Asn Met Asn Phe Val Arg His Leu Val Met Phe Ser Ser Leu Ala Gly 340 345 350

Tyr His Gly Asn Val Gly Gin Ser Asp Tyr Ala Met Ala Asn Glu Ser 355 360 365Tyr His Gly Asn Val Gly Gin Ser Asp Tyr Ala Met Ala Asn Glu Ser 355 360 365

Leu Asn Lys lie Gly Phe Arg Leu Gly Ala Ala Tyr Ser Gin Leu Cys 370 375 380Leu Asn Lys lie Gly Phe Arg Leu Gly Ala Ala Tyr Ser Gin Leu Cys 370 375 380

Val Lys Ser lie Cys Phe Gly Pro Trp Asp Gly Gly Met Val Thr Pro 385 390 395 400Val Lys Ser lie Cys Phe Gly Pro Trp Asp Gly Gly Met Val Thr Pro 385 390 395 400

Ala Leu Lys Lys Gin Phe Gin Ser Met Gly Val Gin lie lie Pro Arg 405 410 415Ala Leu Lys Lys Gin Phe Gin Ser Met Gly Val Gin lie lie Pro Arg 405 410 415

Glu Gly Gly Ala Glu Thr Val Ala Arg lie Val Leu Ser Ser Asn Pro 420 425 430Glu Gly Gly Ala Glu Thr Val Ala Arg lie Val Leu Ser Ser Asn Pro 420 425 430

Ser Gin Val Leu Val Gly Asn Trp Gly Val Pro Pro Val Ser Pro Leu 435 440 445Ser Gin Val Leu Val Gly Asn Trp Gly Val Pro Pro Val Ser Pro Leu 435 440 445

Ser Lys Ser Ala Thr lie Val Gin Thr Phe Thr Pro Glu Leu Asn Pro 450 455 460 177 201038734 s Ly u e L e 5 h 6 p 4Ser Lys Ser Ala Thr lie Val Gin Thr Phe Thr Pro Glu Leu Asn Pro 450 455 460 177 201038734 s Ly u e L e 5 h 6 p 4

Ser His Gin lie His Gly Lys Asn Val Leu Pro Met Thr 470 475 480Ser His Gin lie His Gly Lys Asn Val Leu Pro Met Thr 470 475 480

Val Ala lie Gly Tyr Leu Ala His Leu Val Lys Asn Phe Tyr Ala Gly 485 490 495 His His Leu Trp Gly Val Glu Asp Ala Gin Leu Phe Ser Gly Val Val 500 505 510 lie Asp His Ala Val Gin Ala Gin Val Lys Leu Thr Glu Gin Ser Leu 515 520 525 Asp Asp Asp Gly Lys Val Lys Val Gin Ala Val Leu Thr Ala Ser Asn 530 535 540 Asp Asn Gly Lys Met Val Pro Ala Tyr Lys Ala Val lie Val Leu Gly 545 550 555 560Val Ala lie Gly Tyr Leu Ala His Leu Val Lys Asn Phe Tyr Ala Gly 485 490 495 His His Leu Trp Gly Val Glu Asp Ala Gin Leu Phe Ser Gly Val Val 500 505 510 lie Asp His Ala Val Gin Ala Gin Val Lys Leu Thr Glu Gin Ser Leu 515 520 525 Asp Asp Asp Gly Lys Val Lys Val Gin Ala Val Leu Thr Ala Ser Asn 530 535 540 Asp Asn Gly Lys Met Val Pro Ala Tyr Lys Ala Val lie Val Leu Gly 545 550 555 560

Lys Thr Ser Arg Pro Ala Phe lie Leu Lys Asp Phe Ser Leu Gin Glu 565 570 575Lys Thr Ser Arg Pro Ala Phe lie Leu Lys Asp Phe Ser Leu Gin Glu 565 570 575

Ser Asn Ser Arg Ser Ala Asp Glu Leu Tyr Asp Gly Lys 580 585 e h p u e L r o h 9 T 5Ser Asn Ser Arg Ser Ala Asp Glu Leu Tyr Asp Gly Lys 580 585 e h p u e L r o h 9 T 5

u e L 〇5 r 9 p 5 y Tx G s •1 Hu e L 〇5 r 9 p 5 y Tx G s •1 H

Phe Arg Gly lie Thr Lys Leu Leu Asn Val Ser Asp 600 605Phe Arg Gly lie Thr Lys Leu Leu Asn Val Ser Asp 600 605

Thr Ser Leu Thr Thr Gin Cys Thr Asn 工le Asp Leu Thr Ala Thr Glu 610 615 620Thr Ser Leu Thr Thr Gin Cys Thr Asn worker Le Asp Leu Thr Ala Thr Glu 610 615 620

Arg Gly Gin Phe Ala Asp lie Glu Pro Val Asn Pro Phe Met Ala Asp 625 630 635 640Arg Gly Gin Phe Ala Asp lie Glu Pro Val Asn Pro Phe Met Ala Asp 625 630 635 640

Ala Ala Phe Gin Ala Met Leu Val Trp Val Arg Asn Leu Arg Asn Ser 645 650 655Ala Ala Phe Gin Ala Met Leu Val Trp Val Arg Asn Leu Arg Asn Ser 645 650 655

A n s A 〇o r 6 p 6 u e L r e sA n s A 〇o r 6 p 6 u e L r e s

Asn Cys Glu Arg Val Asp lie Tyr Lys Pro lie 665 670Asn Cys Glu Arg Val Asp lie Tyr Lys Pro lie 665 670

Ala Pro Gly Glu Lys Tyr Tyr Thr Thr Leu Gin Ala Leu Gly Asn Thr 675 680 685Ala Pro Gly Glu Lys Tyr Tyr Thr Thr Leu Gin Ala Leu Gly Asn Thr 675 680 685

a V r e s yo 19 G 6 r e sa V r e s yo 19 G 6 r e s

a V r 5 e 9 s 6 s Ly u e La V r 5 e 9 s 6 s Ly u e L

Phe Tyr Met His Asp Glu Gin Gly 700Phe Tyr Met His Asp Glu Gin Gly 700

Glu Val Phe Leu Ser Gly Arg Ala 705 710 a V 1 5 3 i—I V 7 _~I a V r e sGlu Val Phe Leu Ser Gly Arg Ala 705 710 a V 1 5 3 i—I V 7 _~I a V r e s

n s A &lt;21〇&gt; 102 &lt;211〉 1311 &lt;212&gt; DNA &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 102 ttaagtgttg gtgataatat ttgtcatgat caacgtgttg ctgttgttgg tatggctgtt 60 atgtatgctg gttgtcaaaa tcaacatgaa ttttggcaat ctttacaagg taaaaatatg 120 aattcaaaat cgatttcaca aaatcgttta ggttctgagt atagagaaga acattttaaa 180 178 240 240Ns A &lt;21〇&gt; 102 &lt;211> 1311 &lt;212&gt; DNA &lt;213&gt; Thraustochytrium species &lt;400&gt; 102 ttaagtgttg gtgataatat ttgtcatgat caacgtgttg ctgttgttgg tatggctgtt 60 atgtatgctg gttgtcaaaa tcaacatgaa ttttggcaat ctttacaagg taaaaatatg 120 aattcaaaat cgatttcaca aaatcgttta Ggttctgagt atagagaaga acattttaaa 180 178 240 240

Ο 201038734 cctgaaagaa gtaaatattc cgataccttt tgtaatgaaa gatatggttg tattgatgag aatgttcaaa gtgaacatga acttttatta aaacttgcaa aagatgctat tgcggataca aaaggttcta ttgatttgaa taaaaccgga atcgttagtg gttgcttatc ttttccaatg gataatttac aaggtgattt attaaatttg tatcaatgtc acattgaaaa gaaaattggg ccaaatgcat taaaagatgt gaatttatgg tctaaaagaa ccaccaacgg aaaagatgat aaaaaagctt attttgatcc tgcctctttc gtagctgaac aattagatat gggaccatta cattatagtt tagatgctgc ttgtgcgtct gcactttatg tattaagact tgctcaagat catttattaa gtggtgctgc tgatacaatg ttatgtggtg catcttgttt acctgaacct ttttttattt tatctggttt ttctactttt catgcaatgc cattatctgg tgatgtttct gctcctttgc ataaaacttc acaaggtctt acacctggtg aaggtggtgc tattatggta cttaaacgat taaatgatgc aatccgtgat ggtgatagaa tttatggtac tttacttggt gctgaattaa gtaatgctgg ttgtggttta ccattgagtc cacatatgcc aagtgaattt gattgtatgg aaaaagcttt acaaagagta cacagattac catcatctat tcaatatgtt gagtgtcatg caactggtac accacaaggt gataaagttg aaattgatgc tatgacaaaa tgttttggtg aacatttacc aaggtttggt tcaacgaaag ggaattttgg tcatacactt gttgctgctg gttttgctgg tatgtgtaaa gttttattat caatgcaata tggtgaaata ccaccaactc caggtcttga aaatccagac aatattatgc atgatttagt tgttactgaa acaattccat ggcctaatac aaatggtgat ttgaaacgtg catgtttatc tgcttttgga ttcggtggta ctaatgcaca tgctgtattt gaagagtatc gttcagattt a &lt;210&gt; 103 &lt;211&gt; 437 &lt;212&gt; PRT &lt;213〉 破囊壺菌物種 &lt;400&gt; 103Ο 201038734 cctgaaagaa gtaaatattc cgataccttt tgtaatgaaa gatatggttg tattgatgag aatgttcaaa gtgaacatga acttttatta aaacttgcaa aagatgctat tgcggataca aaaggttcta ttgatttgaa taaaaccgga atcgttagtg gttgcttatc ttttccaatg gataatttac aaggtgattt attaaatttg tatcaatgtc acattgaaaa gaaaattggg ccaaatgcat taaaagatgt gaatttatgg tctaaaagaa ccaccaacgg aaaagatgat aaaaaagctt attttgatcc tgcctctttc gtagctgaac aattagatat gggaccatta cattatagtt tagatgctgc ttgtgcgtct gcactttatg tattaagact tgctcaagat catttattaa gtggtgctgc tgatacaatg ttatgtggtg catcttgttt acctgaacct ttttttattt tatctggttt ttctactttt catgcaatgc cattatctgg tgatgtttct gctcctttgc ataaaacttc acaaggtctt acacctggtg aaggtggtgc tattatggta cttaaacgat taaatgatgc aatccgtgat ggtgatagaa tttatggtac tttacttggt gctgaattaa gtaatgctgg ttgtggttta ccattgagtc cacatatgcc aagtgaattt gattgtatgg aaaaagcttt acaaagagta cacagattac catcatctat tcaatatgtt gagtgtcatg caactggtac accacaaggt gataaagttg aaattgatgc tatgacaaaa tgttttggtg aacatttacc aaggtttggt tcaacgaaag ggaattttgg tcatacac tt gttgctgctg gttttgctgg tatgtgtaaa gttttattat caatgcaata tggtgaaata ccaccaactc caggtcttga aaatccagac aatattatgc atgatttagt tgttactgaa acaattccat ggcctaatac aaatggtgat ttgaaacgtg catgtttatc tgcttttgga ttcggtggta ctaatgcaca tgctgtattt gaagagtatc gttcagattt a &lt; 210 &gt; 103 &lt; 211 &gt; 437 &lt; 212 &gt; PRT &lt; 213> Thraustochytrium species &lt;400&gt; 103

Leu Ser Val Gly Asp Asn lie Cys His Asp Gin Arg Val Ala Val Val 15 10 15Leu Ser Val Gly Asp Asn lie Cys His Asp Gin Arg Val Ala Val Val 15 10 15

Gly Met Ala Val Met Tyr Ala Gly Cys Gin Asn Gin His Glu Phe Trp 20 25 30Gly Met Ala Val Met Tyr Ala Gly Cys Gin Asn Gin His Glu Phe Trp 20 25 30

Gin Ser Leu Gin Gly Lys Asn Met Asn Ser Lys Ser lie Ser Gin Asn 35 40 45Gin Ser Leu Gin Gly Lys Asn Met Asn Ser Lys Ser lie Ser Gin Asn 35 40 45

Arg Leu Gly Ser Glu Tyr Arg Glu Glu His Phe Lys Pro Glu Arg Ser 50 55 60Arg Leu Gly Ser Glu Tyr Arg Glu Glu His Phe Lys Pro Glu Arg Ser 50 55 60

Lys Tyr Ser Asp Thr Phe Cys Asn Glu Arg Tyr Gly Cys 工le Asp Glu 65 70 75 80Lys Tyr Ser Asp Thr Phe Cys Asn Glu Arg Tyr Gly Cys work le Asp Glu 65 70 75 80

Asn Val Gin Ser Glu His Glu Leu Leu Leu Lys Leu Ala Lys Asp Ala 85 90 95 lie Ala Asp Thr Lys Gly Ser lie Asp Leu Asn Lys Thr Gly lie Val 100 105 110 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1311 179 201038734Asn Val Gin Ser Glu His Glu Leu Leu Leu Lys Leu Ala Lys Asp Ala 85 90 95 lie Ala Asp Thr Lys Gly Ser lie Asp Leu Asn Lys Thr Gly lie Val 100 105 110 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1311 179 201038734

Ser Gly Cys Leu Ser Phe Pro Met Asp Asn Leu Gin Gly Asp Leu Leu 115 120 125Ser Gly Cys Leu Ser Phe Pro Met Asp Asn Leu Gin Gly Asp Leu Leu 115 120 125

Asn Leu Tyr Gin Cys His lie Glu Lys Lys lie Gly Pro Asn Ala Leu 130 135 140Asn Leu Tyr Gin Cys His lie Glu Lys Lys lie Gly Pro Asn Ala Leu 130 135 140

Lys Asp Val Asn Leu Trp Ser Lys Arg Thr Thr Asn Gly Lys Asp Asp 145 150 155 160Lys Asp Val Asn Leu Trp Ser Lys Arg Thr Thr Asn Gly Lys Asp Asp 145 150 155 160

Lys Lys Ala Tyr Phe Asp Pro Ala Ser Phe Val Ala Glu Gin Leu Asp 165 170 175Lys Lys Ala Tyr Phe Asp Pro Ala Ser Phe Val Ala Glu Gin Leu Asp 165 170 175

Met Gly Pro Leu His Tyr Ser Leu Asp Ala Ala Cys Ala Ser Ala Leu 180 185 190Met Gly Pro Leu His Tyr Ser Leu Asp Ala Ala Cys Ala Ser Ala Leu 180 185 190

Tyr Val Leu Arg Leu Ala Gin Asp His Leu Leu Ser Gly Ala Ala Asp 195 200 205Tyr Val Leu Arg Leu Ala Gin Asp His Leu Leu Ser Gly Ala Ala Asp 195 200 205

Thr Met Leu Cys Gly Ala Ser Cys Leu Pro Glu Pro Phe Phe lie Leu 210 215 220Thr Met Leu Cys Gly Ala Ser Cys Leu Pro Glu Pro Phe Phe lie Leu 210 215 220

Ser Gly Phe Ser Thr Phe His Ala Met Pro Leu Ser Gly Asp Val Ser 225 230 235 240Ser Gly Phe Ser Thr Phe His Ala Met Pro Leu Ser Gly Asp Val Ser 225 230 235 240

Ala Pro Leu His Lys Thr Ser Gin Gly Leu Thr Pro Gly Glu Gly Gly 245 250 255Ala Pro Leu His Lys Thr Ser Gin Gly Leu Thr Pro Gly Glu Gly Gly 245 250 255

Ala lie Met Val Leu Lys Arg Leu Asn Asp Ala lie Arg Asp Gly Asp 260 265 270Ala lie Met Val Leu Lys Arg Leu Asn Asp Ala lie Arg Asp Gly Asp 260 265 270

Arg lie Tyr Gly Thr Leu Leu Gly Ala Glu Leu Ser Asn Ala Gly Cys 275 280 285Arg lie Tyr Gly Thr Leu Leu Gly Ala Glu Leu Ser Asn Ala Gly Cys 275 280 285

Gly Leu Pro Leu Ser Pro His Met Pro Ser Glu Phe Asp Cys Met Glu 290 295 300Gly Leu Pro Leu Ser Pro His Met Pro Ser Glu Phe Asp Cys Met Glu 290 295 300

Lys Ala Leu Gin Arg Val His Arg Leu Pro Ser Ser lie Gin Tyr Val 305 310 315 320Lys Ala Leu Gin Arg Val His Arg Leu Pro Ser Ser lie Gin Tyr Val 305 310 315 320

Glu Cys His Ala Thr Gly Thr Pro Gin Gly Asp Lys Val Glu lie Asp 325 330 335Glu Cys His Ala Thr Gly Thr Pro Gin Gly Asp Lys Val Glu lie Asp 325 330 335

Ala Met Thr Lys Cys Phe Gly Glu His Leu Pro Arg Phe Gly Ser Thr 340 345 350Ala Met Thr Lys Cys Phe Gly Glu His Leu Pro Arg Phe Gly Ser Thr 340 345 350

Lys Gly Asn Phe Gly His Thr Leu Val Ala Ala Gly Phe Ala Gly Met 355 360 365Lys Gly Asn Phe Gly His Thr Leu Val Ala Ala Gly Phe Ala Gly Met 355 360 365

Cys Lys Val Leu Leu Ser Met Gin Tyr Gly Glu lie Pro Pro Thr Pro 370 375 380Cys Lys Val Leu Leu Ser Met Gin Tyr Gly Glu lie Pro Pro Thr Pro 370 375 380

Gly Leu Glu Asn Pro Asp Asn lie Met His Asp Leu Val Val Thr Glu 385 390 395 400Gly Leu Glu Asn Pro Asp Asn lie Met His Asp Leu Val Val Thr Glu 385 390 395 400

Thr lie Pro Trp Pro Asn Thr Asn Gly Asp Leu Lys Arg Ala Cys Leu 405 410 415 180 201038734Thr lie Pro Trp Pro Asn Thr Asn Gly Asp Leu Lys Arg Ala Cys Leu 405 410 415 180 201038734

Ser Ala Phe Gly 420Ser Ala Phe Gly 420

Phe Gly Gly Thr Asa Ala His Ala Val Phe Glu Glu 425 430Phe Gly Gly Thr Asa Ala His Ala Val Phe Glu Glu 425 430

Tyr Arg Ser Asp Leu 435Tyr Arg Ser Asp Leu 435

&lt;210&gt; 104 &lt;211&gt; 1323 &lt;212&gt; DNA &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 104 aaaattgcta gaacgtgcta tttcttggag tgttatatta gatattctaa tctggtgtta ttatatcgtc gatattcctt tatacatctt ggtccatcat agatggttct ggtagtgctg ccatttgcaa ttggttttga gattcaattg tccatagcag tctggtagaa atacaaagag ggtgcagcaa ccaaattgga cattctagag tgcgaaaatg teg ttgttggtat tttacaatgg aagataaaga aagaagtgga gacctttaca aaccaaatgg ategtgeteg tacttgaaaa atattggaaa tcacgattac tagetaatgg aaaatctttt attttgaatc aaegattgag ctgttggtga egaaagatat tcacttgtga ttgcaattgg gtttaatcaa ataagccaac catggttgaa gttcgtgtta ggaatctgaa tggtcatggt atttttacaa aactgatttt gttgttagct caaagttgca tgttgcatta gttaatgaac tttggttgca tgaaggtgaa tgaagtagat tgtaaaatct aaatgctgat tgattgtacg tgaagttggc tgaactggca agatgaacta tagtgtgaaa aacggctttg gaaagatgtt aaatgttgat tggaatcgta tttggtactt gcatgtgatt gcttgtggtt aaaaggttac gtttcgatta gttttagttg aaggaaegee tatgtcaatg actcgagttt aattccgtat gctgttgttg cgtagaagta ggatattttg gattcaactg ccaactatta gagctatcag aatgaactgg gctaatgttg tgcctgtaca gaatggtcca gaaaatagac atgtctgatg tgaaaggatt tacctgaaaa tacaaaaatt gtttaccaat tcgacagagc gattaggtac tccaaactgc atagaggtac catcattatg atcgttgtct ttgccggggt aagtttccac ctggagatgg aaaaaattta aacaagcttt caagttcagg gtgaaatttt gagatgttgg accgatattt aatcattttt atgctgtcgt tacaaggaca acaagaattt tagatggaga accaagaggt gatacaggag aettaaegea tgatcttgaa ggtcaaagaa aagtacatca gggttttact tgatttggga tgatttatgt acaaaatgaa ttgtggagct tgcaacggtg gaagaatgea caaacatcat caatgaaggt atatgeatet accaaagtta tgtatgtgaa ttctggagtt tcatgaagaa 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1323 &lt;210&gt; 105 &lt;211&gt; 441 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 105&Lt; 210 &gt; 104 &lt; 211 &gt; 1323 &lt; 212 &gt; DNA &lt; 213 &gt; Thraustochytrium species &lt; 400 &gt; 104 aaaattgcta gaacgtgcta tttcttggag tgttatatta gatattctaa tctggtgtta ttatatcgtc gatattcctt tatacatctt ggtccatcat agatggttct ggtagtgctg ccatttgcaa ttggttttga gattcaattg tccatagcag tctggtagaa atacaaagag ggtgcagcaa ccaaattgga cattctagag tgcgaaaatg teg ttgttggtat tttacaatgg aagataaaga aagaagtgga gacctttaca aaccaaatgg ategtgeteg tacttgaaaa atattggaaa tcacgattac tagetaatgg aaaatctttt attttgaatc aaegattgag ctgttggtga egaaagatat tcacttgtga ttgcaattgg gtttaatcaa ataagccaac catggttgaa gttcgtgtta ggaatctgaa tggtcatggt atttttacaa aactgatttt gttgttagct caaagttgca tgttgcatta gttaatgaac tttggttgca tgaaggtgaa tgaagtagat tgtaaaatct aaatgctgat tgattgtacg tgaagttggc tgaactggca agatgaacta tagtgtgaaa aacggctttg gaaagatgtt aaatgttgat tggaatcgta tttggtactt gcatgtgatt gcttgtggtt aaaaggttac Gtttcgatta gttttagttg aaggaaegee tatgtcaatg actcgagttt aattccgtat gctgttgttg cgtagaagta ggatattttg gattcaactg ccaactatta gagctatca g aatgaactgg gctaatgttg tgcctgtaca gaatggtcca gaaaatagac atgtctgatg tgaaaggatt tacctgaaaa tacaaaaatt gtttaccaat tcgacagagc gattaggtac tccaaactgc atagaggtac catcattatg atcgttgtct ttgccggggt aagtttccac ctggagatgg aaaaaattta tgtatgtgaa ttctggagtt aacaagcttt caagttcagg gtgaaatttt gagatgttgg accgatattt aatcattttt atgctgtcgt tacaaggaca acaagaattt tagatggaga accaagaggt gatacaggag aettaaegea tgatcttgaa ggtcaaagaa aagtacatca gggttttact tgatttggga tgatttatgt acaaaatgaa ttgtggagct tgcaacggtg gaagaatgea caaacatcat caatgaaggt atatgeatet accaaagtta Tcatgaagaa 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1323 &lt;210&gt; 105 &lt;211&gt; 441 &lt;212&gt; PRT &lt;213&gt; Thraustochytrium species&lt;400&gt; 105

Lys lie Ala lie Val Gly Met Glu Ser Glu Phe Gly Thr Leu Lys Gly 15 10 15Lys lie Ala lie Val Gly Met Glu Ser Glu Phe Gly Thr Leu Lys Gly 15 10 15

Leu Gin Glu Phe Glu Arg Ala lie Tyr Asn Gly Gly His Gly Ala Cys 20 25 30 181 201038734Leu Gin Glu Phe Glu Arg Ala lie Tyr Asn Gly Gly His Gly Ala Cys 20 25 30 181 201038734

Asp Leu Pro Glu Asn Arg Trp Arg Phe Leu Gly Glu Asp Lys Glu Phe 35 40 45Asp Leu Pro Glu Asn Arg Trp Arg Phe Leu Gly Glu Asp Lys Glu Phe 35 40 45

Leu Gin Ala Cys Gly Leu Gin Lys Leu Pro Arg Gly Cys Tyr lie Lys 50 55 60Leu Gin Ala Cys Gly Leu Gin Lys Leu Pro Arg Gly Cys Tyr lie Lys 50 55 60

Glu Val Glu Thr Asp Phe Lys Arg Leu Arg Leu Pro Met lie Gin Glu 65 70 Ί5 80Glu Val Glu Thr Asp Phe Lys Arg Leu Arg Leu Pro Met lie Gin Glu 65 70 Ί5 80

Asp lie Leu Arg Pro Leu Gin Leu Leu Ala Val Ser lie lie Asp Arg 85 90 95Asp lie Leu Arg Pro Leu Gin Leu Leu Ala Val Ser lie lie Asp Arg 85 90 95

Ala Leu Asn Ala Ser Gly Val Lys Pro Asn Gly Lys Val Ala Val Leu 100 105 110Ala Leu Asn Ala Ser Gly Val Lys Pro Asn Gly Lys Val Ala Val Leu 100 105 110

Val Gly Leu Gly Thr Asp Leu Glu Leu Tyr Arg His Arg Ala Arg Val 115 120 125Val Gly Leu Gly Thr Asp Leu Glu Leu Tyr Arg His Arg Ala Arg Val 115 120 125

Ala Leu Lys Glu Arg Leu Gin Thr Ala Val Lys Glu Asp lie Pro Leu 130 135 140Ala Leu Lys Glu Arg Leu Gin Thr Ala Val Lys Glu Asp lie Pro Leu 130 135 140

Leu Glu Lys Leu Met Asn Tyr Val Asn Asp Arg Gly Thr Ser Thr Ser 145 150 155 160Leu Glu Lys Leu Met Asn Tyr Val Asn Asp Arg Gly Thr Ser Thr Ser 145 150 155 160

Tyr Thr Ser Tyr lie Gly Asn Leu Val Ala Thr Arg Val Ser Ser Leu 165 170 175Tyr Thr Ser Tyr lie Gly Asn Leu Val Ala Thr Arg Val Ser Ser Leu 165 170 175

Trp Gly Phe Thr Gly Pro Ser Phe Thr lie Thr Glu Gly Glu Asn Ser 180 185 190Trp Gly Phe Thr Gly Pro Ser Phe Thr lie Thr Glu Gly Glu Asn Ser 180 185 190

Val Tyr Arg Cys Leu Asp Leu Gly Arg Trp Phe Leu Ala Asn Gly Glu 195 200 205Val Tyr Arg Cys Leu Asp Leu Gly Arg Trp Phe Leu Ala Asn Gly Glu 195 200 205

Val Asp Ala Val Val Val Ala Gly Val Asp Leu Cys Gly Ser Ala Glu 210 215 220Val Asp Ala Val Val Val Ala Gly Val Asp Leu Cys Gly Ser Ala Glu 210 215 220

Asn Leu Phe Val Lys Ser Arg Arg Ser Lys Val Ser Thr Gin Asn Glu 225 230 235 240Asn Leu Phe Val Lys Ser Arg Arg Ser Lys Val Ser Thr Gin Asn Glu 225 230 235 240

Pro Phe Ala Asn Phe Glu Ser Asn Ala Asp Gly Tyr Phe Ala Gly Asp 245 250 255Pro Phe Ala Asn Phe Glu Ser Asn Ala Asp Gly Tyr Phe Ala Gly Asp 245 250 255

Gly Cys Gly Ala Leu Val Leu Lys Arg Leu Ser Asp Cys Thr Asp Ser 260 265 270Gly Cys Gly Ala Leu Val Leu Lys Arg Leu Ser Asp Cys Thr Asp Ser 260 265 270

Thr Glu Lys lie Tyr Ala Thr Val Asp Ser 工le Ala Val Gly Asp Glu 275 280 285Thr Glu Lys lie Tyr Ala Thr Val Asp Ser work le Ala Val Gly Asp Glu 275 280 285

Val Gly Pro Thr lie Lys Gin Ala Leu Lys Asn Ala Ser lie Ala Ala 290 295 300Val Gly Pro Thr lie Lys Gin Ala Leu Lys Asn Ala Ser lie Ala Ala 290 295 300

Lys Asp lie Glu Leu Ala Glu Leu Ser Ala Ser Ser Gly Lys His His 305 310 315 320Lys Asp lie Glu Leu Ala Glu Leu Ser Ala Ser Ser Gly Lys His His 305 310 315 320

Ser Gly Arg lie Thr Cys Glu Asp Glu Leu Asn Glu Leu Gly Glu lie 182 201038734 325 330 335Ser Gly Arg lie Thr Cys Glu Asp Glu Leu Asn Glu Leu Gly Glu lie 182 201038734 325 330 335

Phe Asn Glu Gly lie Gin Arg Val Ala lie Gly Ser Val Lys Ala Asn 340 345 350Phe Asn Glu Gly lie Gin Arg Val Ala lie Gly Ser Val Lys Ala Asn 340 345 350

Val Gly Asp Val Gly Tyr Ala Ser Gly Ala Ala Ser Leu lie Lys Thr 355 360 365Val Gly Asp Val Gly Tyr Ala Ser Gly Ala Ala Ser Leu lie Lys Thr 355 360 365

Ala Leu Cys Leu Tyr Asn Arg Tyr Leu Pro Lys Leu Pro Asn Trp Asn 370 375 380Ala Leu Cys Leu Tyr Asn Arg Tyr Leu Pro Lys Leu Pro Asn Trp Asn 370 375 380

Lys Pro Thr Lys Asp Val Glu Trp Ser Lys 385 390 a V e h p e h p r 5 e 9 S3Lys Pro Thr Lys Asp Val Glu Trp Ser Lys 385 390 a V e h p e h p r 5 e 9 S3

u o 1 o G 4 s CYu o 1 o G 4 s CY

His Ser Arg Ala Trp Leu Lys Asn Val Asp Glu Asn Arg His Ala Val 405 410 415His Ser Arg Ala Trp Leu Lys Asn Val Asp Glu Asn Arg His Ala Val 405 410 415

O ❹O ❹

Val Ser Gly Val Cys Glu Asn Gly Ser Cys Tyr Gly lie Val Met Ser 420 425 430Val Ser Gly Val Cys Glu Asn Gly Ser Cys Tyr Gly lie Val Met Ser 420 425 430

Asp Val Gin Gly His His Glu Glu Ser 435 440 &lt;210&gt; 106 &lt;211&gt; 1353 &lt;212&gt; DNA &lt;213〉 破囊壺菌物種 &lt;400&gt; 106 cctacaatag atattgactc caatgtgttt gctgagatgc ttaatctacc gcaggataaa 60 aacaaaaaat ttgcggtcgc attggttacc acaccaaata aactccagcg tgaaatagaa 120 cttgctgtga agggtattcc acgttgcgta aaagcaaaaa gagattggtg ttctccatct 180 ggaagtattt ttgcttgtaa tccactcaaa agtgataata ttgcatttat gtatggtgaa 240 ggccgaagcc catatgctgg actgggatat gatttgcatc gaatttggcc tatgctacac 300 gagttggtta acaatagaac tacagaactt tgggatcaag gtgatagttg gtatttacct 360 cgatctagct ctgttgctga aaaagaaaaa gtcttcggag attttgataa gaatcaaatt 420 gaaatgttta gattgggtat ttttgtatca atgtgtttca ctgatatggc cactgaactt 480 ttgggtttaa aacccaaagc cgcgtttggt ttaagtttgg gtgaaatatc tatgcttttt 540 gcattttcta aaaagaatac caagttgtcc aaagaattga cccgtcgtct aaaagaagca 600 aaagtttggg catcacaatt agctgttgaa tttgcagcta ttcgagattt gtggaatatt 660 ccagctgata aatctattga tgaattttgg caagggtatt ttgtttacgc aaatcgaacc 720 ctggtcgaga acacaattgg ggagaataaa tttgttcgtt tgttgattgt aaatgattcg 780 caaagttgtc taattgccgg gaaaccagat gaatgtcaaa aagttattga gaagcttcat 840 ttgaagctac cggcggttcc agtaactcag ggtatgatcg gtcattgccc agaagcaatt 900 ccttatctag atcaaatcag tcatattcat gaaatgcttg aaattccaaa acccgaaaat 960 gtgaaattgt ttacaactag tgaaaacaga gaattagtgt cgatgaaaga ttccgtgtca 1020 aaattggttg ctgagattta tcagcatgtt gctgattttc caaacatcgt gaacaaggtt 1080 aaagaaactt gcaaaactga tatatttatt gaattgggat cgaacaatta tcgatctgga 1140 183 201038734 gctgtcaaaa caattttagg tccagaaatc gtttctgttg caattgatag gcaaaatgaa 1200 actgcatggg gtcaactaat gaagatggtt gcatcgttga taagtcatcg agttccgggt 1260 gttgaattga aaaaactcta tcatcctgaa ttgctgaaat ttgatccaca ggcaaaaccg 1320 aatcgtttca tcagaaatat agaactgaat gga 1353 &lt;210&gt; 107 &lt;211&gt; 451 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 107Asp Val Gin Gly His His Glu Glu Ser 435 440 &lt;210&gt; 106 &lt;211&gt; 1353 &lt;212&gt; DNA &lt;213> Thraustochytrium Species &lt;400&gt; 106 cctacaatag atattgactc caatgtgttt gctgagatgc ttaatctacc gcaggataaa 60 aacaaaaaat ttgcggtcgc attggttacc acaccaaata aactccagcg tgaaatagaa 120 cttgctgtga agggtattcc acgttgcgta aaagcaaaaa gagattggtg ttctccatct 180 ggaagtattt ttgcttgtaa tccactcaaa agtgataata ttgcatttat gtatggtgaa 240 ggccgaagcc catatgctgg actgggatat gatttgcatc gaatttggcc tatgctacac 300 gagttggtta acaatagaac tacagaactt tgggatcaag gtgatagttg gtatttacct 360 cgatctagct ctgttgctga aaaagaaaaa gtcttcggag attttgataa gaatcaaatt 420 gaaatgttta gattgggtat ttttgtatca atgtgtttca ctgatatggc cactgaactt 480 ttgggtttaa aacccaaagc cgcgtttggt ttaagtttgg Gtgaaatatc tatgcttttt 540 gcattttcta aaaagaatac caagttgtcc aaagaattga cccgtcgtct aaaagaagca 600 aaagtttggg catcacaatt agctgttgaa tttgcagcta ttcgagattt gtggaatatt 660 ccagctgata aatctattga tgaattttgg caagggtatt ttgtttacgc aaatcgaacc 720 ctggtcgaga acacaattgg ggagaataaa tttgttcgtt tgttgattgt aaatgattcg 780 caaagttgtc taattgccgg gaaaccagat gaatgtcaaa aagttattga gaagcttcat 840 ttgaagctac cggcggttcc agtaactcag ggtatgatcg gtcattgccc agaagcaatt 900 ccttatctag atcaaatcag tcatattcat gaaatgcttg aaattccaaa acccgaaaat 960 gtgaaattgt ttacaactag tgaaaacaga gaattagtgt cgatgaaaga ttccgtgtca 1020 aaattggttg ctgagattta tcagcatgtt gctgattttc caaacatcgt gaacaaggtt 1080 aaagaaactt gcaaaactga tatatttatt gaattgggat cgaacaatta tcgatctgga 1140 183 201038734 gctgtcaaaa caattttagg tccagaaatc gtttctgttg caattgatag gcaaaatgaa 1200 actgcatggg gtcaactaat gaagatggtt gcatcgttga taagtcatcg agttccgggt 1260 gttgaattga aaaaactcta tcatcctgaa ttgctgaaat ttgatccaca ggcaaaaccg 1320 aatcgtttca tcagaaatat agaactgaat gga 1353 &lt; 210 &gt; 107 &lt; 211 &gt; 451 &lt; 212 &gt; PRT &lt; 213 &gt; Thraustochytrium species &lt;400&gt; 107

Pro Thr lie Asp lie Asp Ser Asn Val Phe Ala Glu Met Leu Asn Leu 15 10 15Pro Thr lie Asp lie Asp Ser Asn Val Phe Ala Glu Met Leu Asn Leu 15 10 15

Pro Gin Asp Lys Asn Lys Lys Phe Ala Val Ala Leu Val Thr Thr Pro 20 25 30Pro Gin Asp Lys Asn Lys Lys Phe Ala Val Ala Leu Val Thr Thr Pro 20 25 30

Asn Lys Leu Gin Arg Glu lie Glu Leu Ala Val Lys Gly lie Pro Arg 35 40 45Asn Lys Leu Gin Arg Glu lie Glu Leu Ala Val Lys Gly lie Pro Arg 35 40 45

Cys Val Lys Ala Lys Arg Asp Trp Cys Ser Pro Ser Gly Ser lie Phe 50 55 60Cys Val Lys Ala Lys Arg Asp Trp Cys Ser Pro Ser Gly Ser lie Phe 50 55 60

Ala Cys Asn Pro Leu Lys Ser Asp Asn lie Ala Phe Met Tyr Gly Glu 65 70 75 80Ala Cys Asn Pro Leu Lys Ser Asp Asn lie Ala Phe Met Tyr Gly Glu 65 70 75 80

Gly Arg Ser Pro Tyr Ala Gly Leu Gly Tyr Asp Leu His Arg lie Trp 85 90 95Gly Arg Ser Pro Tyr Ala Gly Leu Gly Tyr Asp Leu His Arg lie Trp 85 90 95

Pro Met Leu His Glu Leu Val Asn Asn Arg Thr Thr Glu Leu Trp Asp 100 105 110Pro Met Leu His Glu Leu Val Asn Asn Arg Thr Thr Glu Leu Trp Asp 100 105 110

Gin Gly Asp Ser Trp Tyr Leu Pro Arg Ser Ser Ser Val Ala Glu Lys 115 120 125Gin Gly Asp Ser Trp Tyr Leu Pro Arg Ser Ser Ser Val Ala Glu Lys 115 120 125

Glu Lys Val Phe Gly Asp Phe Asp Lys Asn Gin lie Glu Met Phe Arg 130 135 140Glu Lys Val Phe Gly Asp Phe Asp Lys Asn Gin lie Glu Met Phe Arg 130 135 140

Leu Gly lie Phe Val Ser Met Cys Phe Thr Asp Met Ala Thr Glu Leu 145 150 155 160Leu Gly lie Phe Val Ser Met Cys Phe Thr Asp Met Ala Thr Glu Leu 145 150 155 160

Leu Gly Leu Lys Pro Lys Ala Ala Phe Gly Leu Ser Leu Gly Glu lie 165 170 175Leu Gly Leu Lys Pro Lys Ala Ala Phe Gly Leu Ser Leu Gly Glu lie 165 170 175

Ser Met Leu Phe Ala Phe Ser Lys Lys Asn Thr Lys Leu Ser Lys Glu 180 185 190Ser Met Leu Phe Ala Phe Ser Lys Lys Asn Thr Lys Leu Ser Lys Glu 180 185 190

Leu Thr Arg Arg Leu Lys Glu Ala Lys Val Trp Ala Ser Gin Leu Ala 195 200 205Leu Thr Arg Arg Leu Lys Glu Ala Lys Val Trp Ala Ser Gin Leu Ala 195 200 205

Val Glu Phe Ala Ala lie Arg Asp Leu Trp Asn lie Pro Ala Asp Lys 210 215 220 r 5 e2 s 2 C：Val Glu Phe Ala Ala lie Arg Asp Leu Trp Asn lie Pro Ala Asp Lys 210 215 220 r 5 e2 s 2 C:

He Asp Glu Phe Trp Gin Gly Tyr Phe Val Tyr Ala Asn Arg Thr 230 235 240 184 201038734He Asp Glu Phe Trp Gin Gly Tyr Phe Val Tyr Ala Asn Arg Thr 230 235 240 184 201038734

Leu Val Glu Asn Thr lie Gly Glu Asn Lys Phe Val Arg Leu Leu lie 245 250 255Leu Val Glu Asn Thr lie Gly Glu Asn Lys Phe Val Arg Leu Leu lie 245 250 255

Val Asn Asp Ser Gin Ser Cys 260Val Asn Asp Ser Gin Ser Cys 260

s Y' c u I—I G po s 7 A 2 0 r p syy _—I G a A e 5 16 1 2 u e Ls Y' c u I—I G po s 7 A 2 0 r p syy _—I G a A e 5 16 1 2 u e L

Gin Lys Val lie Glu Lys Leu His Leu Lys Leu Pro Ala Val Pro Val 275 280 285 Thr Gin Gly Met lie Gly His Cys Pro Glu Ala lie Pro Tyr Leu Asp 290 295 300Gin Lys Val lie Glu Lys Leu His Leu Lys Leu Pro Ala Val Pro Val 275 280 285 Thr Gin Gly Met lie Gly His Cys Pro Glu Ala lie Pro Tyr Leu Asp 290 295 300

Gin lie Ser His 工le His Glu Met Leu Glu 305 310 η o S2 A 3 u •—I G o r p s y L 0 r p e 5 _—_ -—_ 1 3Gin lie Ser His Le His Glu Met Leu Glu 305 310 η o S2 A 3 u •—I G o r p s y L 0 r p e 5 _—_ ——_ 1 3

Val Lys Leu Phe Thr Thr Ser Glu Asn Arg Glu Leu Val 325 330 s y· t 5 e3 M3 r e s oVal Lys Leu Phe Thr Thr Ser Glu Asn Arg Glu Leu Val 325 330 s y· t 5 e3 M3 r e s o

Asp Ser Val Ser Lys Leu Val Ala Glu lie Tyr Gin His Val Ala Asp 340 345 350Asp Ser Val Ser Lys Leu Val Ala Glu lie Tyr Gin His Val Ala Asp 340 345 350

Phe Pro Asn lie Val Asn Lys Val Lys Glu Thr Cys Lys Thr Asp lie 355 360 365 6 o 17 13 e h pPhe Pro Asn lie Val Asn Lys Val Lys Glu Thr Cys Lys Thr Asp lie 355 360 365 6 o 17 13 e h p

Glu Leu Gly Ser Asn Asn Tyr Arg Ser Gly Ala Val Lys Thr 375 380 lie Leu Gly Pro Glu 工le Val Ser Val Ala 工le Asp Arg Gin Asn Glu 385 390 395 400Glu Leu Gly Ser Asn Asn Tyr Arg Ser Gly Ala Val Lys Thr 375 380 lie Leu Gly Pro Glu work le Val Ser Val Ala work as As Arg Gin Asn Glu 385 390 395 400

Thr Ala Trp Gly Gin Leu Met Lys Met Val Ala Ser Leu lie 405 410 s i H r 5 el s 4Thr Ala Trp Gly Gin Leu Met Lys Met Val Ala Ser Leu lie 405 410 s i H r 5 el s 4

Arg Val Pro Gly Val Glu Leu Lys Lys Leu Tyr His Pro Glu Leu Leu 420 425 430 Lys Phe Asp Pro Gin Ala Lys Pro Asn Arg Phe lie Arg Asn lie Glu 435 440 445Arg Val Pro Gly Val Glu Leu Lys Lys Leu Tyr His Pro Glu Leu Leu 420 425 430 Lys Phe Asp Pro Gin Ala Lys Pro Asn Arg Phe lie Arg Asn lie Glu 435 440 445

Leu Asn Gly 450 &lt;210&gt; 108 &lt;211&gt; 1203 &lt;212&gt; DNA &lt;213〉 破囊壺菌物種 &lt;400&gt; 108 ctcaaaacat atgaggttga ctatcctttg tacacaggtg ccatggctaa aggaattgcg 60 tctgctgatt tggttattgc tgctggtaaa tcaaagatct tggcatcatt tggagctggt 120 gggttggcct tacaagtggt agaagatgcc attaaacaaa ttaaagctga attggggaac 180 ggtccgtttg ctgtaaattt gattcattca ccattcgatc ctagcttgga gaagggtaac 240 gttgatcttt ttctaaaata taacgttcga tttgttgaag tatccgcatt tatgtcatta 300 acccctcagg ttgtacgata cagagccgct ggtttggcca aagcaagaga tggatctgtg 360 185 201038734 aaaattcaaa atcgtattat tgccaaaatt tcaagaacag agttagcgga actgttcttg 420 aaaccagcac ccaaaaatat tttagatgca ttggttgcgg atggatctat tagtcaagaa 480 caagcccaac ttgcattact tgtgccaatg gctgatgata ttactgtgga agctgattct 540 ggtgggcata ctgacaatcg accaattcat gttttgttac ctttgataat tcagcaaaga 600 aatagaattt gtaaacaata cccaaaacat ttaaaagttc gaatcggagc agctggtggt 660 attggatgcc cgaaggcagc atttgctgcg tttgagatgg gtgctgcata cattgcaact 720 ggaacggtaa atcaactttc aaaggaagca ggtacttgtg actatgtacg taaagtattg 780 aataaagcta catattcgga tgttaccatg gctccagccg cagatatgtt cgatcatggt 840 gttgaattac aagttttgaa gaaaggtact atgtttcctt cacgtgctaa aaaactatac 900 gatttgttca aaaaatacaa atcgattgag gaattaccag cagatgaggt gaaaaaactt 960 gagcaaaaag ttttcaaaaa gtcgtttgat gaagtatggg atgagaccaa gaattactat 1020 attaatcgtt tacattctcc cgaaaaaatt gaacgtgctg aaagagatgc aaaacttaaa 1080Leu Asn Gly 450 &lt;210&gt; 108 &lt;211&gt; 1203 &lt;212&gt; DNA &lt;213&gt; Thraustochytrium species &lt;400&gt; 108 ctcaaaacat atgaggttga ctatcctttg tacacaggtg ccatggctaa aggaattgcg 60 tctgctgatt tggttattgc tgctggtaaa tcaaagatct tggcatcatt tggagctggt 120 gggttggcct tacaagtggt agaagatgcc attaaacaaa ttaaagctga attggggaac 180 ggtccgtttg ctgtaaattt gattcattca ccattcgatc ctagcttgga gaagggtaac 240 gttgatcttt ttctaaaata taacgttcga tttgttgaag tatccgcatt tatgtcatta 300 acccctcagg ttgtacgata cagagccgct ggtttggcca aagcaagaga tggatctgtg 360 185 201038734 aaaattcaaa atcgtattat tgccaaaatt tcaagaacag agttagcgga actgttcttg 420 aaaccagcac ccaaaaatat tttagatgca ttggttgcgg atggatctat tagtcaagaa 480 caagcccaac ttgcattact tgtgccaatg gctgatgata ttactgtgga agctgattct 540 ggtgggcata ctgacaatcg a aga acc acc acc acc acc acc acc acc acc acttgtg actatgtacg taaagtattg 780 aataaagcta catattcgga tgttaccatg gctccagccg cagatatgtt cgatcatggt 840 gttgaattac aagttttgaa gaaaggtact atgtttcctt cacgtgctaa aaaactatac 900 gatttgttca aaaaatacaa atcgattgag gaattaccag cagatgaggt gaaaaaactt 960 gagcaaaaag ttttcaaaaa gtcgtttgat gaagtatggg atgagaccaa gaattactat 1020 attaatcgtt tacattctcc cgaaaaaatt gaacgtgctg aaagagatgc aaaacttaaa 1080

atgtcgttat gttttcgttg gtatttgtcg aagtcttcca gatgggctaa taccggtgaa 1140 tctggaagag tgcaggatta tcaaatttgg tgtggtccag caattgggtc atataatgat 1200 ttt 1203 &lt;210&gt; 109 &lt;211&gt; 401 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;4〇〇&gt; 109Atgtcgttat gttttcgttg gtatttgtcg aagtcttcca gatgggctaa taccggtgaa 1140 tctggaagag tgcaggatta tcaaatttgg tgtggtccag caattgggtc atataatgat 1200 ttt 1203 &lt;210&gt; 109 &lt;211&gt; 401 &lt;212&gt; PRT &lt;213&gt; Thraustochytrium species &lt;4〇〇&gt; 109

Leu Lys Thr Tyr Glu Val Asp Tyr Pro Leu Tyr Thr Gly Ala Met Ala 15 10 15Leu Lys Thr Tyr Glu Val Asp Tyr Pro Leu Tyr Thr Gly Ala Met Ala 15 10 15

Lys Gly lie Ala Ser Ala Asp Leu Val lie Ala Ala Gly Lys Ser Lys 20 25 30 lie Leu Ala Ser Phe Gly Ala Gly Gly Leu Ala Leu Gin Val Val Glu 35 40 45Lys Gly lie Ala Ser Ala Asp Leu Val lie Ala Ala Gly Lys Ser Lys 20 25 30 lie Leu Ala Ser Phe Gly Ala Gly Gly Leu Ala Leu Gin Val Val Glu 35 40 45

Asp Ala lie Lys Gin lie Lys Ala Glu Leu Gly Asn Gly Pro Phe Ala 50 55 60Asp Ala lie Lys Gin lie Lys Ala Glu Leu Gly Asn Gly Pro Phe Ala 50 55 60

Val Asn Leu lie His Ser Pro Phe Asp Pro Ser Leu Glu Lys Gly Asn 65 70 75 80Val Asn Leu lie His Ser Pro Phe Asp Pro Ser Leu Glu Lys Gly Asn 65 70 75 80

Val Asp Leu Phe Leu Lys Tyr Asn Val Arg Phe Val Glu Val Ser Ala 85 90 95Val Asp Leu Phe Leu Lys Tyr Asn Val Arg Phe Val Glu Val Ser Ala 85 90 95

Phe Met Ser Leu Thr Pro Gin Val Val Arg Tyr Arg Ala Ala Gly Leu 100 105 110Phe Met Ser Leu Thr Pro Gin Val Val Arg Tyr Arg Ala Ala Gly Leu 100 105 110

Ala Lys Ala Arg Asp Gly Ser Val Lys lie Gin Asn Arg lie lie Ala 115 120 125Ala Lys Ala Arg Asp Gly Ser Val Lys lie Gin Asn Arg lie lie Ala 115 120 125

Lys lie Ser Arg Thr Glu Leu Ala Glu Leu Phe Leu Lys Pro Ala Pro 130 135 140Lys lie Ser Arg Thr Glu Leu Ala Glu Leu Phe Leu Lys Pro Ala Pro 130 135 140

Lys Asn lie Leu Asp Ala Leu Val Ala Asp Gly Ser lie Ser Gin Glu 186 201038734 145 150 155 160 Gin Ala Gin Leu Ala 165 Leu Leu Val Pro Met 170 Ala Asp Asp lie Thr 175 ValLys Asn lie Leu Asp Ala Leu Val Ala Asp Gly Ser lie Ser Gin Glu 186 201038734 145 150 155 160 Gin Ala Gin Leu Ala 165 Leu Leu Val Pro Met 170 Ala Asp Asp lie Thr 175 Val

Glu Ala Asp Ser Gly Gly His Thr Asp Asn Arg Pro lie His Val Leu 180 185 190Glu Ala Asp Ser Gly Gly His Thr Asp Asn Arg Pro lie His Val Leu 180 185 190

Leu Pro Leu lie lie Gin Gin Arg Asn Arg lie Cys Lys Gin Tyr Pro 195 200 205Leu Pro Leu lie lie Gin Gin Arg Asn Arg lie Cys Lys Gin Tyr Pro 195 200 205

Lys His Leu Lys Val Arg lie Gly Ala Ala Gly Gly 工le Gly Cys Pro 210 215 220Lys His Leu Lys Val Arg lie Gly Ala Ala Gly Gly work le Gly Cys Pro 210 215 220

Lys Ala Ala Phe Ala Ala Phe Glu Met Gly Ala Ala Tyr lie Ala Thr 225 230 235 240 oLys Ala Ala Phe Ala Ala Phe Glu Met Gly Ala Ala Tyr lie Ala Thr 225 230 235 240 o

Gly Thr Val Asn Gin Leu Ser Lys Glu Ala Gly Thr Cys Asp Tyr Val 245 250 255Gly Thr Val Asn Gin Leu Ser Lys Glu Ala Gly Thr Cys Asp Tyr Val 245 250 255

Arg Lys Val Leu Asn Lys Ala Thr Tyr Ser Asp Val Thr Met Ala Pro 260 265 270Arg Lys Val Leu Asn Lys Ala Thr Tyr Ser Asp Val Thr Met Ala Pro 260 265 270

Ala Ala Asp Met Phe Asp His Gly Val Glu Leu Gin Val Leu Lys Lys 275 280 285Ala Ala Asp Met Phe Asp His Gly Val Glu Leu Gin Val Leu Lys Lys 275 280 285

Gly Thr Met Phe Pro Ser Arg Ala Lys Lys Leu Tyr Asp Leu Phe Lys 290 295 300Gly Thr Met Phe Pro Ser Arg Ala Lys Lys Leu Tyr Asp Leu Phe Lys 290 295 300

Lys Tyr Lys Ser lie Glu Glu Leu Pro Ala Asp Glu Val Lys Lys Leu 305 310 315 320Lys Tyr Lys Ser lie Glu Glu Leu Pro Ala Asp Glu Val Lys Lys Leu 305 310 315 320

Glu Gin Lys Val Phe Lys Lys Ser Phe Asp Glu Val Trp Asp Glu Thr 325 330 335Glu Gin Lys Val Phe Lys Lys Ser Phe Asp Glu Val Trp Asp Glu Thr 325 330 335

Lys Asn Tyr Tyr 工le Asn Arg Leu His Ser Pro Glu Lys lie Glu Arg 340 345 350Lys Asn Tyr Tyr work le Asn Arg Leu His Ser Pro Glu Lys lie Glu Arg 340 345 350

Ala Glu Arg Asp Ala Lys Leu Lys Met Ser Leu Cys Phe Arg Trp Tyr 355 360 365Ala Glu Arg Asp Ala Lys Leu Lys Met Ser Leu Cys Phe Arg Trp Tyr 355 360 365

Leu Ser Lys Ser Ser Arg Trp Ala Asn Thr Gly Glu Ser Gly Arg Val 370 375 380Leu Ser Lys Ser Ser Arg Trp Ala Asn Thr Gly Glu Ser Gly Arg Val 370 375 380

Gin Asp Tyr Gin lie Trp Cys Gly Pro Ala lie Gly Ser Tyr Asn Asp 385 390 395 400Gin Asp Tyr Gin lie Trp Cys Gly Pro Ala lie Gly Ser Tyr Asn Asp 385 390 395 400

Phe &lt;210〉 110 &lt;211&gt; 1350 &lt;212&gt; DNA &lt;213〉 破袭壺菌物種 &lt;400&gt; 110 atggttggtt tacaaatgaa aaagaaacca gtatgggaga tgagtaagga agaacaaagt 187 60 201038734 tctggaaaga ggtaaagtct cctgcgagag ttcagagttg caaggtggtg atatcttata actacgttga gtaactggat tgttatgtga gatgaagaac aaatctattg atgttttctg agtggacttc attactaaaa attcttgaac ggctcacttg ttacatgatg cgttgccgtg gaaatgggat gatgtaaact ggtaatttgt accgtgaaat atgttgtatt ttggacctaa aatatcttct gatctagaat atgttccatg tgggtattga cgttttacgg ttgcaaaagg atggacgatt ttgcagcagg ttccaaaatc aaaatgatat aaggtattga tacaacataa gtgatcattg ttagtgatgg tggttccaga gacaaattag tcaatgaatc atgaacttgg caaagaaaat catcaaatat tgactatgat gtttgatatt agttaccaga ggttactgaa ggctgttctt ttttcaatgt tgttgctcat tatgcacggt attaatcgaa taaaggagtt cattaaacct ggaaaaattg ctacaagtta tggtggtgca gtattttcca ttgcagtcag ttttcaattt tccacatcgt aactggacaa tcaatcattt tgtggttgat cattgattct gaattgttgg atcgataagt gttactttga tatgatgttc gttgaatctg aaaggtgatc gagggtgaaa gaaatctcca atgagagatg attaaaactg tttgctctaa tgtgagcgtc tgtgcacgga tatggtcttg tgccattttg ctactaaaac cgtccagttc ggtaaacttg ccatatgcta gatatggctg tttaaaggaa aatttgctga atagtcgacg tggatgctga cagtaaatgg gacaatgtga gtgtctatcg cactagtata tgtttttttt gttgtgcggg ttgctgaact atccagcagt aatgggaaaa aaatgcttat gattattggt taaaggatca tttatatgtt ctggacaacc tttatgttat ttgctgatgt atattgatag ttgctttgca aggtgatatt tgtacgttta agttgggaat tgaactttca tcttatgtta attattaaat cgatattcgt tgaatatgat attttttact tcataaaaga acacaaaaca tgtattgggt gattgatcgt tggcgaaaaa agttatggct atggttgggt aaataaagtt ggaaataaga tgatattatt ttatggacgt aatggaaggt 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1350Phe &lt;210> 110 &lt;211&gt; 1350 &lt;212&gt; DNA &lt;213> Spoiled chytrid species &lt;400&gt; 110 atggttggtt tacaaatgaa aaagaaacca gtatgggaga tgagtaagga agaacaaagt 187 60 201038734 tctggaaaga ggtaaagtct cctgcgagag ttcagagttg caaggtggtg atatcttata actacgttga gtaactggat tgttatgtga gatgaagaac aaatctattg atgttttctg agtggacttc attactaaaa attcttgaac ggctcacttg ttacatgatg cgttgccgtg gaaatgggat gatgtaaact ggtaatttgt accgtgaaat atgttgtatt ttggacctaa aatatcttct gatctagaat atgttccatg tgggtattga cgttttacgg ttgcaaaagg atggacgatt ttgcagcagg ttccaaaatc aaaatgatat aaggtattga tacaacataa gtgatcattg ttagtgatgg tggttccaga gacaaattag tcaatgaatc atgaacttgg caaagaaaat catcaaatat tgactatgat gtttgatatt agttaccaga ggttactgaa ggctgttctt ttttcaatgt tgttgctcat tatgcacggt attaatcgaa taaaggagtt cattaaacct ggaaaaattg ctacaagtta tggtggtgca gtattttcca ttgcagtcag ttttcaattt Tccacatcgt aactggacaa tcaatcattt tgtggttgat cattgattct gaattgttgg atcgataagt gttactttga tatgatgttc gttgaatctg aaaggtgatc gagggtgaaa gaaatctcca atgag agatg attaaaactg tttgctctaa tgtgagcgtc tgtgcacgga tatggtcttg tgccattttg ctactaaaac cgtccagttc ggtaaacttg ccatatgcta gatatggctg tttaaaggaa aatttgctga atagtcgacg tggatgctga cagtaaatgg gacaatgtga gtgtctatcg cactagtata tgtttttttt gttgtgcggg ttgctgaact atccagcagt aatgggaaaa aaatgcttat gattattggt taaaggatca tttatatgtt ctggacaacc tttatgttat ttgctgatgt atattgatag ttgctttgca aggtgatatt tgtacgttta agttgggaat tgaactttca tcttatgtta attattaaat cgatattcgt tgaatatgat attttttact tcataaaaga acacaaaaca tgtattgggt gattgatcgt tggcgaaaaa agttatggct atggttgggt Aaataaagtt ggaaataaga tgatattatt ttatggacgt aatggaaggt 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1350

&lt;210&gt; 111 &lt;211&gt; 450 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 u &lt;400&gt; 111&lt;210&gt; 111 &lt;211&gt; 450 &lt;212&gt; PRT &lt;213&gt; Thraustochytrium species u &lt;400&gt;

Met Val Gly Leu Gin Met Lys Lys Lys Pro Val Trp Glu Met Ser Lys 15 10 15Met Val Gly Leu Gin Met Lys Lys Lys Pro Val Trp Glu Met Ser Lys 15 10 15

Glu Glu Gin Ser Ser Gly Lys Asn Val Val Phe Asp Tyr Asp Glu Leu 20 25 30Glu Glu Gin Ser Ser Gly Lys Asn Val Val Phe Asp Tyr Asp Glu Leu 20 25 30

Leu Glu Phe Ala Glu Gly Asp lie Gly Lys Val Phe Gly Pro Lys Phe 35 40 45Leu Glu Phe Ala Glu Gly Asp lie Gly Lys Val Phe Gly Pro Lys Phe 35 40 45

Asp lie lie Asp Lys Tyr Ser Arg Arg Val Arg Leu Pro Ala Arg Glu 50 55 60Asp lie lie Asp Lys Tyr Ser Arg Arg Val Arg Leu Pro Ala Arg Glu 50 55 60

Tyr Leu Leu Val Thr Arg Val Thr Leu Met Asp Ala Glu Val Gly Asn 65 70 75 80Tyr Leu Leu Val Thr Arg Val Thr Leu Met Asp Ala Glu Val Gly Asn 65 70 75 80

Phe Arg Val Gly Ser Arg Met Val Thr Glu Tyr Asp Val Pro Val Asn 85 90 95 188 201038734Phe Arg Val Gly Ser Arg Met Val Thr Glu Tyr Asp Val Pro Val Asn 85 90 95 188 201038734

Gly Glu Leu Ser Gin Gly Gly Asp Val Pro Trp Ala Val Leu Val Glu 100 105 110Gly Glu Leu Ser Gin Gly Gly Asp Val Pro Trp Ala Val Leu Val Glu 100 105 110

Ser Gly Gin Cys Asp Leu Met Leu lie Ser Tyr Met Gly lie Asp Phe 115 120 125Ser Gly Gin Cys Asp Leu Met Leu lie Ser Tyr Met Gly lie Asp Phe 115 120 125

Gin Cys Lys Gly Asp Arg Val Tyr Arg Leu Leu Asn Thr Thr Leu Thr 130 135 140Gin Cys Lys Gly Asp Arg Val Tyr Arg Leu Leu Asn Thr Thr Leu Thr 130 135 140

Phe Tyr Gly Val Ala His Glu Gly Glu Thr Leu Val Tyr Asp lie Arg 145 150 155 160Phe Tyr Gly Val Ala His Glu Gly Glu Thr Leu Val Tyr Asp lie Arg 145 150 155 160

Val Thr Gly Phe Ala Lys Gly Met His Gly Glu lie Ser Met Phe Phe 165 170 175Val Thr Gly Phe Ala Lys Gly Met His Gly Glu lie Ser Met Phe Phe 165 170 175

Phe Glu Tyr Asp Cys Tyr Val Asn Gly Arg Leu Leu lie Glu Met Arg 180 185 190 ΟPhe Glu Tyr Asp Cys Tyr Val Asn Gly Arg Leu Leu lie Glu Met Arg 180 185 190 Ο

Asp Gly Cys Ala Gly Phe Phe Thr Asp Glu Glu Leu Ala Ala Gly Lys 195 200 205Asp Gly Cys Ala Gly Phe Phe Thr Asp Glu Glu Leu Ala Ala Gly Lys 195 200 205

Gly Val lie Lys Thr Val Ala Glu Leu His Lys Arg Lys Ser 工le Val 210 215 220Gly Val lie Lys Thr Val Ala Glu Leu His Lys Arg Lys Ser Le Val 210 215 220

Pro Lys Ser lie Lys Pro Phe Ala Leu Asn Pro Ala Val His Lys Thr 225 230 235 240Pro Lys Ser lie Lys Pro Phe Ala Leu Asn Pro Ala Val His Lys Thr 225 230 235 240

Met Phe Ser Glu Asn Asp Met Glu Lys Leu Cys Glu Arg Gin Trp Glu 245 250 255Met Phe Ser Glu Asn Asp Met Glu Lys Leu Cys Glu Arg Gin Trp Glu 245 250 255

Asn Val Leu Gly Ser Gly Leu Gin Gly lie Asp Tyr Lys Leu Cys Ala 260 265 270Asn Val Leu Gly Ser Gly Leu Gin Gly lie Asp Tyr Lys Leu Cys Ala 260 265 270

Arg Lys Met Leu Met lie Asp Arg lie Thr Lys lie Gin His Asn Gly 275 280 285Arg Lys Met Leu Met lie Asp Arg lie Thr Lys lie Gin His Asn Gly 275 280 285

Gly Ala Tyr Gly Leu Gly Leu Leu Val Gly Glu Lys lie Leu Glu Arg 290 295 300Gly Ala Tyr Gly Leu Gly Leu Leu Val Gly Glu Lys lie Leu Glu Arg 290 295 300

Asp His Trp Tyr Phe Pro Cys His Phe Val Lys Asp Gin Val Met Ala 305 310 315 320Asp His Trp Tyr Phe Pro Cys His Phe Val Lys Asp Gin Val Met Ala 305 310 315 320

Gly Ser Leu Val Ser Asp Gly Cys Ser Gin Leu Leu Lys Leu Tyr Met 325 330 335Gly Ser Leu Val Ser Asp Gly Cys Ser Gin Leu Leu Lys Leu Tyr Met 325 330 335

Leu Trp Leu Gly Leu His Asp Val Val Pro Asp Phe Gin Phe Arg Pro 340 345 350Leu Trp Leu Gly Leu His Asp Val Val Pro Asp Phe Gin Phe Arg Pro 340 345 350

Val Pro Gly Gin Pro Asn Lys Val Arg Cys Arg Gly Gin lie Ser Pro 355 360 365Val Pro Gly Gin Pro Asn Lys Val Arg Cys Arg Gly Gin lie Ser Pro 355 360 365

His Arg Gly Lys Leu Val Tyr Val Met Glu lie Arg Glu Met Gly Phe 370 375 380His Arg Gly Lys Leu Val Tyr Val Met Glu lie Arg Glu Met Gly Phe 370 375 380

Asn Glu Ser Thr Gly Gin Pro Tyr Ala lie Ala Asp Val Asp lie lie 385 390 395 400 189 201038734Asn Glu Ser Thr Gly Gin Pro Tyr Ala lie Ala Asp Val Asp lie lie 385 390 395 400 189 201038734

Asp Val Asn Tyr Glu Leu Gly Gin Ser Phe Asp Met Ala Asp lie Asp 405 410 415Asp Val Asn Tyr Glu Leu Gly Gin Ser Phe Asp Met Ala Asp lie Asp 405 410 415

Ser Tyr Gly Arg Gly Asn Leu Ser Lys Lys lie Val Val Asp Phe Lys 420 425 430Ser Tyr Gly Arg Gly Asn Leu Ser Lys Lys lie Val Val Asp Phe Lys 420 425 430

Gly lie Ala Leu Gin Met Glu Gly Thr Val Lys Ser Ser Asn lie lie 435 440 445Gly lie Ala Leu Gin Met Glu Gly Thr Val Lys Ser Ser Asn lie lie 435 440 445

r o e 5 s 4 p s A &lt;210&gt; 112 &lt;211&gt; 1200 &lt;212&gt; DNA &lt;213&gt; 破囊壺菌物種r o e 5 s 4 p s A &lt;210&gt; 112 &lt;211&gt; 1200 &lt;212&gt; DNA &lt;213&gt; Thraustochytrium species

O &lt;400&gt; 112 tgcttcaaac catttcctgg taatccttta gataacgatc atacacctgg taaaatgcct 60 ttaacatggt ttaatatgtc cgagtttatg tgtggtaaag tatcaaattg tcttggacca 120 gaatttaaga gatttgataa ctctaaaaca tccagaagtc ctgcctttga tcttgcactt 180 gttacacgtg ttgtgagtgt atcagatatg gaatttaaac ctcatttaaa tattgatgtt 240 aatccaagta agggtacaat gataggtgaa tttgattgcc ctgcagatgc gtggtttttt 300 caaggatcat gtaacgatgg tcatatgccg tattctattg ttatggaiaat tgctcttcaa 360 acttctggtg tattaacttc agttttgaaa gcacctttga ctatggataa agatgatatt 420 cttttccgca atttggatgc cactgctgaa atggttcgaa gtgatgttga ttgtagaggt 480 aaaactatca aaaactttac tcaatgtacc ggttacagta tgctcggaaa aatgggaatt 540 catagattca catttgaatt atctgttgat gatgtagttt tctacaaagg atcaacatct 600 tttggttggt tcacccctga agtattcgag tcacaagttg gtcttgataa tggtaaaaaa 660 gtacaaccat ggtatttgga acaaaaatca tctaatgtag taacttatga cgttgcgtcc 720 actgctggca aggataagtt attttcaaag attggatcta aggatgcaca agttcaaaga 780 agaaatacac aatgtgagtt tctagatact atgcatatta ttccaaatac tggaaagtac 840 aacaaaggtt atgctcatgg agaaaagaaa gttaatccaa acgactggtt cttttcctgt 900 catttctggt ttgatcctgt gatgcctggt tcattaggta ttgaaagtat gtttcaactc 960 attgaagcat tttcaattga tcaaggaatc gcttcaaaac atggtattgt gaatccaact 1020 tttgctcatt ccaatggaaa aacttcttgg aaatacagag gtcaattgaa taacaaaggt 1080 aaacgaatgg atagtgaaat tcatatcaaa gatattgtca aaaatgctga tggtactgtt 1140 gatttgattg ctgatggatt tttattggtt gattcactaa gagtatactc tgcagatgat 1200 &lt;210&gt; &lt;211&gt; &lt;212&gt; &lt;213&gt; 113 400O &lt; 400 &gt; 112 tgcttcaaac catttcctgg taatccttta gataacgatc atacacctgg taaaatgcct 60 ttaacatggt ttaatatgtc cgagtttatg tgtggtaaag tatcaaattg tcttggacca 120 gaatttaaga gatttgataa ctctaaaaca tccagaagtc ctgcctttga tcttgcactt 180 gttacacgtg ttgtgagtgt atcagatatg gaatttaaac ctcatttaaa tattgatgtt 240 aatccaagta agggtacaat gataggtgaa tttgattgcc ctgcagatgc gtggtttttt 300 caaggatcat gtaacgatgg tcatatgccg tattctattg ttatggaiaat tgctcttcaa 360 acttctggtg tattaacttc agttttgaaa gcacctttga ctatggataa agatgatatt 420 cttttccgca atttggatgc cactgctgaa atggttcgaa gtgatgttga ttgtagaggt 480 aaaactatca aaaactttac tcaatgtacc ggttacagta tgctcggaaa aatgggaatt 540 catagattca catttgaatt atctgttgat gatgtagttt tctacaaagg atcaacatct 600 tttggttggt tcacccctga tggtaaaaaa agtattcgag tcacaagttg gtcttgataa agttcaaaga 780 agaaatacac 660 gtacaaccat ggtatttgga acaaaaatca tctaatgtag taacttatga cgttgcgtcc 720 actgctggca aggataagtt attttcaaag attggatcta aggatgcaca aatgtgagtt tctagatact Atgcatatta ttccaaatac tggaaagtac 840 aa caaaggtt atgctcatgg agaaaagaaa gttaatccaa acgactggtt cttttcctgt 900 catttctggt ttgatcctgt gatgcctggt tcattaggta ttgaaagtat gtttcaactc 960 attgaagcat tttcaattga tcaaggaatc gcttcaaaac atggtattgt gaatccaact 1020 tttgctcatt ccaatggaaa aacttcttgg aaatacagag gtcaattgaa taacaaaggt 1080 aaacgaatgg atagtgaaat tcatatcaaa gatattgtca aaaatgctga tggtactgtt 1140 gatttgattg ctgatggatt tttattggtt gattcactaa gagtatactc tgcagatgat 1200 &lt; 210 &gt; &lt; 211 &gt;&lt;212&gt;&lt;213&gt; 113 400

PRT 破囊壺菌物種 &lt;400&gt; 113PRT Thraustochytrium Species &lt;400&gt; 113

Cys Phe Lys Pro Phe Pro Gly Asn Pro Leu Asp Asn Asp His Thr Pro 15 10 15 190 201038734Cys Phe Lys Pro Phe Pro Gly Asn Pro Leu Asp Asn Asp His Thr Pro 15 10 15 190 201038734

Gly Lys Met Pro Leu Thr 20 Trp Phe Asn Met Ser Glu Phe Met Cys Gly 25 30 Lys Val Ser Asn Cys Leu 35 Gly Pro Glu Phe Lys Arg Phe Asp Asn Ser 40 45 Lys Thr Ser Arg Ser Pro 50 Ala Phe Asp Leu Ala Leu Val Thr Arg Val 55 60 Val Ser Val Ser Asp Met 65 70 Glu Phe Lys Pro His Leu Asn lie Asp Val 75 80 Asn Pro Ser Lys Gly Thr 85 Met lie Gly Glu Phe Asp Cys Pro Ala Asp 90 95 Ala Trp Phe Phe Gin Gly 100 Ser Cys Asn Asp Gly His Met Pro Tyr Ser 105 110 O lie Val Met Glu 工le Ala 115 Leu Gin Thr Ser Gly Val Leu Thr Ser Val 120 125 Leu Lys Ala Pro Leu Thr 130 Met Asp Lys Asp Asp lie Leu Phe Arg Asn 135 140 Leu Asp Ala Thr Ala Glu - 145 150 Met Val Arg Ser Asp Val Asp Cys Arg Gly 155 160 - Lys Thr lie Lys Asn Phe 165 Thr Gin Cys Thr Gly Tyr Ser Met Leu Gly 170 175 Lys Met Gly lie His Arg 180 Phe Thr Phe Glu Leu Ser Val Asp Asp Val 185 190 Val Phe Tyr Lys Gly Ser 195 Thr Ser Phe Gly Trp Phe Thr Pro Glu Val 200 205 Phe Glu Ser Gin Val Gly 〇 210 Leu Asp Asn Gly Lys Lys Val Gin Pro Trp 215 220 Tyr Leu Glu Gin Lys Ser 225 230 Ser Asn Val Val Thr Tyr Asp Val Ala Ser 235 240 Thr Ala Gly Lys Asp Lys 245 Leu Phe Ser Lys lie Gly Ser Lys Asp Ala 250 255 Gin Val Gin Arg Arg Asn 260 Thr Gin Cys Glu Phe Leu Asp Thr Met His 265 270 lie lie Pro Asn Thr Gly 275 Lys Tyr Asn Lys Gly Tyr Ala His Gly Glu 280 285 Lys Lys Val Asn Pro Asn 290 Asp Trp Phe Phe Ser Cys His Phe Trp Phe 295 300 Asp Pro Val Met Pro Gly 305 310 Ser Leu Gly lie Glu Ser Met Phe Gin Leu 315 320 191 201038734 r 5 e 2 s 3 e h p a _—_ A u _—I G e lie Asp Gin Gly e _—I I y5 13 G 3 s i H s Y' L r e s a I—_ A β o 1 3 I 3Gly Lys Met Pro Leu Thr 20 Trp Phe Asn Met Ser Glu Phe Met Cys Gly 25 30 Lys Val Ser Asn Cys Leu 35 Gly Pro Glu Phe Lys Arg Phe Asp Asn Ser 40 45 Lys Thr Ser Arg Ser Pro 50 Ala Phe Asp Leu Ala Leu Val Thr Arg Val 55 60 Val Ser Val Ser Asp Met 65 70 Glu Phe Lys Pro His Leu Asn lie Asp Val 75 80 Asn Pro Ser Lys Gly Thr 85 Met lie Gly Glu Phe Asp Cys Pro Ala Asp 90 95 Ala Trp Phe Phe Gin Gly 100 Ser Cys Asn Asp Gly His Met Pro Tyr Ser 105 110 O lie Val Met Glu work le Ala 115 Leu Gin Thr Ser Gly Val Leu Thr Ser Val 120 125 Leu Lys Ala Pro Leu Thr 130 Met Asp Lys Asp Asp lie Leu Phe Arg Asn 135 140 Leu Asp Ala Thr Ala Glu - 145 150 Met Val Arg Ser Asp Val Asp Cys Arg Gly 155 160 - Lys Thr lie Lys Asn Phe 165 Thr Gin Cys Thr Gly Tyr Ser Met Leu Gly 170 175 Lys Met Gly lie His Arg 180 Phe Thr Phe Glu Leu Ser Val Asp Asp Val 185 190 Val Phe Tyr Lys Gly Ser 195 Thr Ser Phe Gly Trp Phe Thr Pro Glu Val 200 205 Phe Glu Ser Gin Val Gly 〇210 Leu Asp Asn Gly Lys Lys Val Gin Pro Trp 215 220 Tyr Leu Glu Gin Lys Ser 225 230 Ser Asn Val Val Thr Tyr Asp Val Ala Ser 235 240 Thr Ala Gly Lys Asp Lys 245 Leu Phe Ser Lys lie Gly Ser Lys Asp Ala 250 255 Gin Val Gin Arg Arg Asn 260 Thr Gin Cys Glu Phe Leu Asp Thr Met His 265 270 lie lie Pro Asn Thr Gly 275 Lys Tyr Asn Lys Gly Tyr Ala His Gly Glu 280 285 Lys Lys Val Asn Pro Asn 290 Asp Trp Phe Phe Ser Cys His Phe Trp Phe 295 300 Asp Pro Val Met Pro Gly 305 310 Ser Leu Gly lie Glu Ser Met Phe Gin Leu 315 320 191 201038734 r 5 e 2 s 3 ehpa ___ A u _—IG e lie Asp Gin Gly e _—II y5 13 G 3 si H s Y' L resa I—_ A β o 1 3 I 3

Val Asn Pro Thr Phe Ala His Ser Asn Gly Lys Thr Ser Trp Lys Tyr 340 345 350 s .1 H e _—_ I u Tx G r 5 e 6 s 3Val Asn Pro Thr Phe Ala His Ser Asn Gly Lys Thr Ser Trp Lys Tyr 340 345 350 s .1 H e ___ I u Tx G r 5 e 6 s 3

Arg Gly Gin Leu Asn Asn Lys Gly Lys Arg Met Asp 355 360 lie Lys Asp lie Val Lys Asn Ala Asp Gly Thr Val Asp Leu lie Ala 370 375 380Arg Gly Gin Leu Asn Asn Lys Gly Lys Arg Met Asp 355 360 lie Lys Asp lie Val Lys Asn Ala Asp Gly Thr Val Asp Leu lie Ala 370 375 380

Asp Gly Phe Leu Leu Val Asp Ser Leu Arg Val Tyr Ser Ala Asp Asp 385 390 395 400 &lt;210&gt; 114 &lt;211&gt; 1729 &lt;212&gt; DNA &lt;213〉 破棄壺菌物種 &lt;400&gt; 114 aggaatcgct tcaaaacatg gtattgtgaa tccaactttt gctcattcca atggaaaaac 60 ttcttggaaa tacagaggtc aattgaataa caaaggtaaa cgaatggata gtgaaattca 120 tatcaaagat attgtcaaaa atgctgatgg tactgttgat ttgattgctg atggattttt 180 attggttgat tcactaagag tatactctgc agatgatctt cgcgtaaaaa ttgtaccggg 240 aaccaaagct gcacctaaat cagtagctgc tgctccaaga catgttgcaa caccaattcc 300 aggagtgcct tcgaatacaa gcagtgttga aatcagtttg gaatctttga agaaagaatt 360 gttaaatctt gagaaaccat tgtatcttga aacttccaat catattgtaa aacaattcgg 420 tgacgttaac aatggccaag catccgttat tccaccatgc accatcaatg atttgggtga 480 gcgtagtttt atggaaacat acaatgttgt tgcaccactt tacactggag ccatggctaa 540 aggtattgca tctgctgatt tggtaattgc agctggtaaa agaaaaattt tgggttcttt 600 tggcgctgga ggcttaccaa tgcacttggt tcgtgcttct gttgaaaaaa tccaagccgc 660 acttccagaa ggtccatacg ctgtcaactt gattcatagt ccattcgact caaatcttga 720 aaagggaaat gtagatctat ttttggaaaa aggtgttcat gttgttgaag catctgcatt 780 cactgctctg accactcaag tagttcgtta ccgtgcatgt ggtttatctc gggctaaaga 840 cggatctgta ttgatcaaaa atagaatcat cggtaaagtt tcaagaaccg aattggctga 900 aatgtttttc agacctgcac cacaaaactt gcttgacaag cttattgcta gtggagaaat 960 cactaaagaa caagcttcat tggctttgga agtaccaatg gctgatgatg tagctgttga 1020 agctgatagc ggtggacata ctgataatag accaattcat gtaatcctac ctttgattat 1080 caatctacga aatagaattc ataaagaatg tggttttcct gctgctttga gagttcgcgt 1140 tggtgctggt ggtggaattg gttgtccaag tgctgcagtt gctgcattca atatgggagc 1200 tgcattcttg attactggca gcgtcaacca agttagcaaa caatctggta cgtgtgatat 1260 cgttagaaag caattatctg aagcttcgta ttcagatatt accatggcac cagcggctga 1320 tatgtttgat caaggagtcg agcttcaagt attaaaaaaa ggaactatgt ttccatctcg 1380 tgcaaagaaa ttgtatgaat tattctgtat gtacaactca tttgatgaca tgccaaaaag 1440 192 1500 201038734 1560 1620 1680 1729 cgaacttcaa agactagaga agcgaatttt tcaaaaatcg cttgcggaag tttgggaaga aactaaagat ttttatatca atcgtttgaa taatcctgag aagattgaac atgctgagaa gaaagatcca aagttgaaga tgtcattatg ctttagatgg tatttgggtt taagttcatt ttgggcaaac aatggaatta aagaaagatc aatggactat caaatttggt gtggtccagc gattggttca tacaatgatt ttgtaaaagg aacttatttg gatcctgca &lt;210&gt; 115 &lt;211&gt; 455 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;40〇&gt; 115Asp Gly Phe Leu Leu Val Asp Ser Leu Arg Val Tyr Ser Ala Asp Asp 385 390 395 400 &lt;210&gt; 114 &lt;211&gt; 1729 &lt;212&gt; DNA &lt;213> Abandoned Species] &lt;400&gt; 114 aggaatcgct tcaaaacatg gtattgtgaa tccaactttt gctcattcca atggaaaaac 60 ttcttggaaa tacagaggtc aattgaataa caaaggtaaa cgaatggata gtgaaattca 120 tatcaaagat attgtcaaaa atgctgatgg tactgttgat ttgattgctg atggattttt 180 attggttgat tcactaagag tatactctgc agatgatctt cgcgtaaaaa ttgtaccggg 240 aaccaaagct gcacctaaat cagtagctgc tgctccaaga catgttgcaa caccaattcc 300 aggagtgcct tcgaatacaa gcagtgttga aatcagtttg gaatctttga agaaagaatt 360 gttaaatctt gagaaaccat tgtatcttga aacttccaat catattgtaa aacaattcgg 420 tgacgttaac aatggccaag catccgttat tccaccatgc accatcaatg atttgggtga 480 gcgtagtttt atggaaacat acaatgttgt tgcaccactt tacactggag ccatggctaa 540 aggtattgca tctgctgatt tggtaattgc agctggtaaa agaaaaattt tgggttcttt 600 tggcgctgga ggcttaccaa tgcacttggt tcgtgcttct gttgaaaaaa tccaagccgc 660 acttccagaa ggtccatacg ctgtcaactt gattcatagt ccattcgact c aaatcttga 720 aaagggaaat gtagatctat ttttggaaaa aggtgttcat gttgttgaag catctgcatt 780 cactgctctg accactcaag tagttcgtta ccgtgcatgt ggtttatctc gggctaaaga 840 cggatctgta ttgatcaaaa atagaatcat cggtaaagtt tcaagaaccg aattggctga 900 aatgtttttc agacctgcac cacaaaactt gcttgacaag cttattgcta gtggagaaat 960 cactaaagaa caagcttcat tggctttgga agtaccaatg gctgatgatg tagctgttga 1020 agctgatagc ggtggacata ctgataatag accaattcat gtaatcctac ctttgattat 1080 caatctacga aatagaattc ataaagaatg tggttttcct gctgctttga gagttcgcgt 1140 tggtgctggt ggtggaattg gttgtccaag tgctgcagtt gctgcattca atatgggagc 1200 tgcattcttg attactggca gcgtcaacca agttagcaaa caatctggta cgtgtgatat 1260 cgttagaaag caattatctg aagcttcgta ttcagatatt accatggcac cagcggctga 1320 tatgtttgat caaggagtcg agcttcaagt attaaaaaaa ggaactatgt ttccatctcg 1380 tgcaaagaaa ttgtatgaat tattctgtat gtacaactca tttgatgaca tgccaaaaag 1440 192 1500 201038734 1560 1620 1680 1729 cgaacttcaa agactagaga agcgaatttt tcaaaaatcg cttgcggaag tttgggaaga aactaaagat Ttttatatca atcgtttgaa taatcctgag aagattgaac atgctgagaa gaaagatcca aagttgaaga tgtcattatg ctttagatgg tatttgggtt taagttcatt ttgggcaaac aatggaatta aagaaagatc aatggactat caaatttggt gtggtccagc gattggttca tacaatgatt ttgtaaaagg aacttatttg gatcctgca &lt; 210 &gt; 115 &lt; 211 &gt; 455 &lt; 212 &gt; PRT &lt; 213 &gt; Thraustochytrium species &lt; 40〇 &gt; 115

Asn Leu Glu Lys Pro Leu Tyr Leu Glu Thr Ser Asn His lie Val Lys 15 10 15Asn Leu Glu Lys Pro Leu Tyr Leu Glu Thr Ser Asn His lie Val Lys 15 10 15

Gin Phe Gly Asp Val Asn Asn O 20Gin Phe Gly Asp Val Asn Asn O 20

Thr lie Asn Asp Leu Gly Glu 35Thr lie Asn Asp Leu Gly Glu 35

Gly Gin Ala Ser Val lie Pro Pro Cys 25 30Gly Gin Ala Ser Val lie Pro Pro Cys 25 30

Arg Ser Phe Met Glu Thr Tyr Asn Val 40 45Arg Ser Phe Met Glu Thr Tyr Asn Val 40 45

Val Ala Pro Leu Tyr Thr Gly 50 55Val Ala Pro Leu Tyr Thr Gly 50 55

Asp Leu Val lie Ala Ala Gly 65 70Asp Leu Val lie Ala Ala Gly 65 70

Ala Met Ala Lys Gly lie Ala Ser Ala 60Ala Met Ala Lys Gly lie Ala Ser Ala 60

Lys Arg Lys lie Leu Gly Ser Phe Gly 75 80Lys Arg Lys lie Leu Gly Ser Phe Gly 75 80

Ala Gly Gly Leu Pro Met His Leu Val Arg Ala Ser Val Glu Lys lie 85 90 95Ala Gly Gly Leu Pro Met His Leu Val Arg Ala Ser Val Glu Lys lie 85 90 95

Gin Ala Ala Leu Pro Glu Gly Pro Tyr Ala Val Asn Leu lie His Ser 100 105 110Gin Ala Ala Leu Pro Glu Gly Pro Tyr Ala Val Asn Leu lie His Ser 100 105 110

Pro Phe Asp Ser Asn Leu Glu Lys Gly Asn Val Asp Leu Phe Leu Glu 115 120 125Pro Phe Asp Ser Asn Leu Glu Lys Gly Asn Val Asp Leu Phe Leu Glu 115 120 125

Lys Gly Val His Val Val Glu Ala Ser Ala Phe Thr Ala Leu Thr Thr 130 135 140Lys Gly Val His Val Val Glu Ala Ser Ala Phe Thr Ala Leu Thr Thr 130 135 140

Gin Val Val Arg Tyr Arg Ala Cys Gly Leu Ser Arg Ala Lys Asp Gly 145 150 155 160Gin Val Val Arg Tyr Arg Ala Cys Gly Leu Ser Arg Ala Lys Asp Gly 145 150 155 160

Ser Val Leu He Lys Asn Arg He He Gly Lys Val Ser Arg Thr Glu 165 170 175Ser Val Leu He Lys Asn Arg He He Gly Lys Val Ser Arg Thr Glu 165 170 175

Leu Ala Glu Met Phe Phe Arg Pro Ala Pro Gin Asn Leu Leu Asp Lys 180 185 190Leu Ala Glu Met Phe Phe Arg Pro Ala Pro Gin Asn Leu Leu Asp Lys 180 185 190

Leu He Ala Ser Gly Glu He Thr Lys Glu Gin Ala Ser Leu Ala Leu 195 200 205Leu He Ala Ser Gly Glu He Thr Lys Glu Gin Ala Ser Leu Ala Leu 195 200 205

Glu Val Pro Met Ala Asp Asp Va! Ala Val Glu Ma Asp Ser GXy Gly 193 201038734 s 5 •12 H 2Glu Val Pro Met Ala Asp Asp Va! Ala Val Glu Ma Asp Ser GXy Gly 193 201038734 s 5 •12 H 2

Thr Asp Asn Arg Pro lie His Val lie Leu Pro Leu lie lie Asn 230 235 240Thr Asp Asn Arg Pro lie His Val lie Leu Pro Leu lie lie Asn 230 235 240

Leu Arg Asn Arg lie His Lys Glu Cys Gly Phe Pro Ala Ala Leu Arg 245 250 255Leu Arg Asn Arg lie His Lys Glu Cys Gly Phe Pro Ala Ala Leu Arg 245 250 255

Val Arg Val Gly Ala Gly Gly Gly lie Gly Cys Pro Ser Ala Ala Val 260 265 270Val Arg Val Gly Ala Gly Gly Gly lie Gly Cys Pro Ser Ala Ala Val 260 265 270

e 5 h 7 p 2 a Γ—I A a _—_ Ae 5 h 7 p 2 a Γ—I A a _—_ A

Asn Met Gly Ala Ala Phe Leu lie Thr Gly Ser Val Asn 280 285Asn Met Gly Ala Ala Phe Leu lie Thr Gly Ser Val Asn 280 285

Gin Val Ser Lys Gin Ser Gly Thr Cys Asp lie Val Arg Lys Gin Leu 290 295 300Gin Val Ser Lys Gin Ser Gly Thr Cys Asp lie Val Arg Lys Gin Leu 290 295 300

Ser Glu Ala Ser Tyr Ser Asp lie Thr Met Ala Pro Ala Ala Asp Met 305 310 315 320Ser Glu Ala Ser Tyr Ser Asp lie Thr Met Ala Pro Ala Ala Asp Met 305 310 315 320

Phe Asp Gin Gly Val Glu Leu Gin Val Leu Lys Lys Gly Thr 325 330 e h p 15 6 3 M3Phe Asp Gin Gly Val Glu Leu Gin Val Leu Lys Lys Gly Thr 325 330 e h p 15 6 3 M3

Pro Ser Arg Ala Lys Lys Leu Tyr Glu Leu Phe Cys Met Tyr Asn Ser 340 345 350Pro Ser Arg Ala Lys Lys Leu Tyr Glu Leu Phe Cys Met Tyr Asn Ser 340 345 350

Phe Asp Asp Met Pro Lys Ser Glu Leu Gin Arg Leu Glu Lys Arg lie 355 360 365Phe Asp Asp Met Pro Lys Ser Glu Leu Gin Arg Leu Glu Lys Arg lie 355 360 365

Phe Gin Lys Ser Leu Ala Glu Val Trp Glu Glu Thr Lys Asp Phe Tyr 370 375 380Phe Gin Lys Ser Leu Ala Glu Val Trp Glu Glu Thr Lys Asp Phe Tyr 370 375 380

s LY u 1 G a _—I A s i H u 5 Ί~ G 3 e 1 Is LY u 1 G a _—I A s i H u 5 Ί~ G 3 e 1 I

He Asn Arg Leu Asn Asn Pro Glu Lys 385 390He Asn Arg Leu Asn Asn Pro Glu Lys 385 390

Asp Pro Lys Leu Lys Met Ser Leu Cys Phe Arg Trp Tyr Leu Gly Leu 405 410 415Asp Pro Lys Leu Lys Met Ser Leu Cys Phe Arg Trp Tyr Leu Gly Leu 405 410 415

Ser Ser Phe Trp Ala Asn Asn Gly lie Lys Glu Arg Ser Met Asp Tyr 420 425 430Ser Ser Phe Trp Ala Asn Asn Gly lie Lys Glu Arg Ser Met Asp Tyr 420 425 430

Gin lie Trp Cys Gly Pro Ala lie Gly Ser Tyr Asn Asp Phe Val Lys 435 440 445Gin lie Trp Cys Gly Pro Ala lie Gly Ser Tyr Asn Asp Phe Val Lys 435 440 445

Gly Thr Tyr Leu Asp Pro Ala 450 455 &lt;210&gt; 116 &lt;211&gt; 903 壺菌物種 &lt;212&gt; DNA &lt;213〉 破囊 &lt;400&gt; 116 tccattcggg aatctggtta cacgattagc ggagaaagat tcacaactga agctcacaaa 60 ttggttactg gaaagcctca tgctccgatt aagaagaagg atgctttcct agtatctggt 120 ggtgctcgtg gtattactcc actttgtatt cgtgaaattg ctaaagcagt gaaaggtggc 180 acttacattt tgatgggtcg atcagctttg gctgatgaac ccttgtgggc taatggtaaa 240 tccggaaaag atttagataa agctggtttg gcatttttga aggaagagtt tgcagctggg 300 194 360 201038734 cgtggtagta aaccaactcc aaaagttcac aaatctttga ttgataaagt gctcggtatt agggaggtta gagcatctat tgcaaatata gaagcccatg gagcaaaagc tatatatttg tcttgcgatg tatcttccgc tgagaaagta aaggctgcag tgcaaaaagt tgaaaaggag catctagttc gtattactgg tattgtgcat gcatcaggcg ttttgaggga taaattggtt gagaacaaaa ctttggatga tttcaacgca gtatatggaa ccaaagtaac tggactagta aacttgctgt cagcagtgaa catgaatttt gttcgtcatt tggttatgtt tagttctttg gctggatatc atggaaatgt tggtcaatct gattatgcaa tggctaacga atcacttaac aagattggtt ttagattggg tgcagcttat tctcaattgt gtgttaaatc tatttgtttt ggaccttggg atggtggaat ggtaactcca gctttgaaaa aacaatttca atcaatgggt gtccagatta ttcctcgtga aggtggcgcg gagactgttg caagaatagt cttatcttca aat &lt;210&gt; 117 &lt;211&gt; 201 &lt;212&gt; PRT &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 117Gly Thr Tyr Leu Asp Pro Ala 450 455 &lt;210&gt; 116 &lt;211&gt; 903 chytrid species &lt;212&gt; DNA &lt;213&gt; rupture &lt;400&gt; 116 tccattcggg aatctggtta cacgattagc ggagaaagat tcacaactga agctcacaaa 60 ttggttactg gaaagcctca tgctccgatt aagaagaagg atgctttcct agtatctggt 120 ggtgctcgtg gtattactcc actttgtatt cgtgaaattg ctaaagcagt gaaaggtggc 180 acttacattt tgatgggtcg atcagctttg gctgatgaac ccttgtgggc taatggtaaa 240 tccggaaaag atttagataa agctggtttg gcatttttga aggaagagtt tgcagctggg 300 194 360 201038734 cgtggtagta aaccaactcc aaaagttcac aaatctttga ttgataaagt gctcggtatt agggaggtta gagcatctat tgcaaatata gaagcccatg gagcaaaagc tatatatttg tcttgcgatg tatcttccgc tgagaaagta aaggctgcag tgcaaaaagt tgaaaaggag catctagttc gtattactgg tattgtgcat gcatcaggcg ttttgaggga taaattggtt Gagaacaaaa ctttggatga tttcaacgca gtatatggaa ccaaagtaac tggactagta aacttgctgt cagcagtgaa catgaatttt gttcgtcatt tggttatgtt tagttctttg gctggatatc atggaaatgt tggtcaatct gattatgcaa tggctaacga atcacttaac aagattggtt ttagattggg tgcagcttat tctcaat Tgt gtgttaaatc tatttgtttt ggaccttggg atggtggaat ggtaactcca gctttgaaaa aacaatttca atcaatgggt gtccagatta ttcctcgtga aggtggcgcg gagactgttg caagaatagt cttatcttca aat &lt;210&gt; 117 &lt;211&gt; 201 &lt;212&gt; PRT &lt;213&gt; Thraustochytrium species &lt;400&gt;

Ο 420 480 540 600 660 720 780 840 900 903Ο 420 480 540 600 660 720 780 840 900 903

Ser 工le Arg Glu Ser Gly Tyr Thr lie Ser Gly Glu Arg Phe Thr Thr 15 10 15Ser worker le Arg Glu Ser Gly Tyr Thr lie Ser Gly Glu Arg Phe Thr Thr 15 10 15

Glu Ala His Lys Leu Val Thr Gly Lys Pro His Ala Pro lie Lys Lys 20 25 30Glu Ala His Lys Leu Val Thr Gly Lys Pro His Ala Pro lie Lys Lys 20 25 30

Lys Asp Ala Phe Leu Val Ser Gly Gly Ala Arg Gly lie Thr Pro Leu 35 40 45Lys Asp Ala Phe Leu Val Ser Gly Gly Ala Arg Gly lie Thr Pro Leu 35 40 45

Cys 工le Arg Glu lie Ala Lys Ala Val Lys Gly Gly Thr Tyr lie Leu 50 55 60Cys work le Arg Glu lie Ala Lys Ala Val Lys Gly Gly Thr Tyr lie Leu 50 55 60

Met Gly Arg Ser Ala Leu Ala Asp Glu Pro Leu Trp Ala Asn Gly Lys 65 70 75 80Met Gly Arg Ser Ala Leu Ala Asp Glu Pro Leu Trp Ala Asn Gly Lys 65 70 75 80

Ser Gly Lys Asp Leu Asp Lys Ala Gly Leu Ala Phe Leu Lys Glu Glu 85 90 95Ser Gly Lys Asp Leu Asp Lys Ala Gly Leu Ala Phe Leu Lys Glu Glu 85 90 95

Phe Ala Ala Gly Arg Gly Ser Lys Pro Thr Pro Lys Val His Lys Ser 100 105 110Phe Ala Ala Gly Arg Gly Ser Lys Pro Thr Pro Lys Val His Lys Ser 100 105 110

Leu lie Asp Lys Val Leu Gly lie Arg Glu Val Arg Ala Ser lie Ala 115 120 125Leu lie Asp Lys Val Leu Gly lie Arg Glu Val Arg Ala Ser lie Ala 115 120 125

Asn lie Glu Ala His Gly Ala Lys Ala lie Tyr Leu Ser Cys Asp Val 130 135 140Asn lie Glu Ala His Gly Ala Lys Ala lie Tyr Leu Ser Cys Asp Val 130 135 140

Ser Ser Ala Glu Lys Val Lys Ala Ala Val Gin Lys Val Glu Lys Glu 145 150 155 160Ser Ser Ala Glu Lys Val Lys Ala Ala Val Gin Lys Val Glu Lys Glu 145 150 155 160

His Leu Val Arg lie Thr Gly lie Val His Ala Ser Gly Val Leu Arg 165 170 175 195 201038734His Leu Val Arg lie Thr Gly lie Val His Ala Ser Gly Val Leu Arg 165 170 175 195 201038734

Asp Lys Leu Val Glu Asn Lys Thr Leu Asp Asp Phe Asn Ala Val Tyr 180 185 190Asp Lys Leu Val Glu Asn Lys Thr Leu Asp Asp Phe Asn Ala Val Tyr 180 185 190

Gly Thr a V s 5 y9 L· τ—_Gly Thr a V s 5 y9 L· τ—_

Thr Gly Leu Val Asn 200 &lt;21〇&gt; 118 &lt;211&gt; 39 &lt;212&gt; DNA &lt;213&gt; 破囊壺菌物種 &lt;400&gt; 118 ctaaagtctc atcaaattca tggtaaaaat gttttgcct 39 &lt;210&gt; &lt;211&gt; &lt;212&gt; &lt;213&gt; 119 13 PRT破囊壺菌物種 &lt;400&gt; 119 Leu Lys Ser His Gin lie His Gly Lys Asn Val Leu Pro 15 10Thr Gly Leu Val Asn 200 &lt;21〇&gt; 118 &lt;211&gt; 39 &lt;212&gt; DNA &lt;213&gt; Thraustochytrium species &lt;400&gt; 118 ctaaagtctc atcaaattca tggtaaaaat gttttgcct 39 &lt;210&gt;&lt;211&gt;&lt;212&gt;&lt;213&gt; 119 13 PRT Thraustochytrium Species &lt;400&gt; 119 Leu Lys Ser His Gin lie His Gly Lys Asn Val Leu Pro 15 10

O &lt;210&gt; &lt;211&gt; &lt;212&gt; &lt;213&gt; &lt;220&gt; &lt;223&gt; 120 8976 DMA人工序列 密碼子最佳化的PFA1 &lt;400&gt; 120 atggaggacc agcgtattgc gatcgttggc cttagcgcga tccttccctc gggcgagaac 60 gtccgcgagt cgtgggaggc gatccgtgac ggcctcaact gcctttccga cctgcccgcc 120 gaccgcgttg acgtcactgc ctactacaac cccacgaagg gcgtcaagga caagatctac 180 tgcaagcgtg gtggcttcat ccccgagtac gagtttgact cgcgcgagtt cggcctcaac 240 atgcttcaga tggaggactc ggacgccaac cagaccctca ccctgctcaa ggttaaggag 300 gccctcgacg acgccaacat tcccgcgttt accaacgaga agaagaacat cggttgcgtc 360 ctcggtattg gcggtggtca gaaggcctcg catgagttct acagccgcct caactacgtc 420 gtcgtggata aggtcctccg caagatgggc ctcccggacg aggacgtcga gactgctgtc 480 gagaagttca aggccaactt tcccgagtgg cgccttgact ccttccccgg ctttctcggt 540 aacgtcactg cgggccgctg caccaacacc ttcaacatgg agggcatgaa ctgcgtggtc 600 gatgccgcct gcgcctcgtc cctcatcgct atcaaggtcg ccatcgatga gctgctccac 660 ggcgattgcg acgcgatgat tgctggcgcg acgtgcaccg acaacgccct tggcatgtac 720 atggcctttt ccaagacccc cgtcttttcc acggaccaga gctgcctcgc ctacgacgag 780 aaaaccaagg gtatgctcat tggcgagggt tccgccatgt tcgtccttaa gcgctacgcc 840 gacgccgtcc gcgatggcga caccgtccac gccgtcatcc gctcgtgctc gtcctcctcc 900 gacggcaagg cgtcgggtat ctacaccccg accatctcgg gccaggagga ggccatcctt 960 cgcgcctacc gtcgtgccgg cgtgagcccg aacacgatca cccttgtgga gggccatggc 1020 accggcaccc ccgtcggcga caagatcgag ctgaccgccc tccgcaacgt ctttgacaag 1080 gcctacggcc ctggccacaa ggaggaggtc gctgtgggct ccatcaagtc gcaqatcggt 1140 196 201038734O &lt;210&gt;&lt;211&gt;&lt;212&gt;&lt;213&gt;&lt;220&gt;&lt;223&gt; 120 8976 DMA artificial sequence codon optimization PFA1 &lt;400&gt; 120 atggaggacc agcgtattgc gatcgttggc cttagcgcga tccttccctc gggcgagaac 60 gtccgcgagt cgtgggaggc gatccgtgac ggcctcaact gcctttccga cctgcccgcc 120 gaccgcgttg acgtcactgc ctactacaac cccacgaagg gcgtcaagga caagatctac 180 tgcaagcgtg gtggcttcat ccccgagtac gagtttgact cgcgcgagtt cggcctcaac 240 atgcttcaga tggaggactc ggacgccaac cagaccctca ccctgctcaa ggttaaggag 300 gccctcgacg acgccaacat tcccgcgttt accaacgaga agaagaacat cggttgcgtc 360 ctcggtattg gcggtggtca gaaggcctcg catgagttct acagccgcct caactacgtc 420 gtcgtggata aggtcctccg caagatgggc ctcccggacg aggacgtcga gactgctgtc 480 gagaagttca aggccaactt tcccgagtgg cgccttgact ccttccccgg ctttctcggt 540 aacgtcactg cgggccgctg caccaacacc ttcaacatgg agggcatgaa ctgcgtggtc 600 gatgccgcct gcgcctcgtc cctcatcgct atcaaggtcg ccatcgatga gctgctccac 660 ggcgattgcg acgcgatgat tgctggcgcg acgtgcaccg acaacgccct tggcatgtac 720 atggcctttt ccaagaccc c cgtcttttcc acggaccaga gctgcctcgc ctacgacgag 780 aaaaccaagg gtatgctcat tggcgagggt tccgccatgt tcgtccttaa gcgctacgcc 840 gacgccgtcc gcgatggcga caccgtccac gccgtcatcc gctcgtgctc gtcctcctcc 900 gacggcaagg cgtcgggtat ctacaccccg accatctcgg gccaggagga ggccatcctt 960 cgcgcctacc gtcgtgccgg cgtgagcccg aacacgatca cccttgtgga gggccatggc 1020 accggcaccc ccgtcggcga caagatcgag ctgaccgccc tccgcaacgt ctttgacaag 1080 gcctacggcc ctggccacaa ggaggaggtc gctgtgggct ccatcaagtc gcaqatcggt 1140 196 201038734

cacctcaagg aagacgctcc atctcggaca gtcccccgtc cttgaggagt atgctcctcc ctcctcaagg tccaagtttc ggctttgctt ttccagaagg agcaccgccc cagtacaccc aacgatatgg caggtcatgt agcaagaccg ttccgcgacg gccctttacg gccatggcta ggtccgaacg tcgccctcgc ctcaagacgc cacatggaga cccaccggct aagacggccc atgtacgccg ctcgtcaacg tccgccaagg gtcgctctca cctcgcaaga caggagcgcg aacacgaaca gttgtccgcg gagcttcaga aagcacaagg tactccagct cccgtgcagg aaggcggagc atcgagctgg ccgtcgccgg cgcagtccat ccacgctcta gtgcgggcat tcgagcccga acgccgtcaa agtcccgcga aggactcctt ccaagagcat acatcacgac tcatcaacga acatgttcaa agaaggccca ttccccgcaa agtactcgca ccggctttgc ccgctggcct tgcgcgacgc cctcgtccat agaccgtcat agggtttccg acgccgagaa cgtcccccaa tctcccgcca ctggcgcccg agattttccc actcggacat ccgactttga aaacgaccct agcgtatcat cgggcgagct tccaggccct aggctcaggc ccatcctcct tttccaaggg ctgccgcacc aggtcgtgct acatggagct ctgcgctggc caacgtcgag catcaacacc ctccagcttc gcagaccaag cgcgaactcg gaagtgcgtc tasgctcaag cgaggacacg caccagctgg caacaagtcc cgacgtcgcg ggaggaggtg gccctacgcg gacgacgacc cccggctttt catcgaccgc ccccaagaag caagctctcg cactggtgcc cgtggtccac gcagtttcag gattttcagc catgactagc cgttttcatc gggcgacacc ccagctccgc taagtgggag caccctctcc gaacgacggt tgagaagctc tgccacccag caccaagtcg cgctatgctg tcaggttgcc ggttcaggtc tgaggtcctc tgagactgag ctcgtcaagc aacccgccca atgaaccgcc ggctttggcg ccctaccgcc ctccagaagc aacacgaaga ggctccgtcc atttccattc gcgctcccga gtggccgccc atgcagtggc atcaacgaca cgcgagagcc gtcgcctcgt gttgcgggcc gaggacctct tccgctgacg gctcccgagg aacagcggcg ctcgcctgcg aaggccctct aacgtcaccg tcggtccagt gagttcggcc agcgtcctca caggcggccg cttaaggacc gccgctacct cgcactgtca aagaagcagc gccagcgccg tcgaacgctg gaggagctgg tcccctgcga agcgcctccg gcctccaaga cttggtatcg tcgtgatggc acctcgtcga cgtggatcac gcgctaacta tgaacgtttc tctgcgagga acaccgacta ccagccagca ttagcgccat aggagggcgc tgttctcggg cgcagttccg agtcggttaa ccctcgacaa ccgtcggcct actcgctcgg ttaagctcgt gcgccatggc tttgggtcgc tccaggccga acggcgcgtt cggccgtcaa gcggtgtctt ttctcaccca ccaagcaggt ctgttagcgt tgcagatggc cgacccgcat acgttagcaa gctgcgtgca tccaggacaa accttcagaa cctcggacgc agactggcaa ccgttcgtgt tggactcggg ccggctacga attcgatcaa tctcaagcat tggcactgtc caagccgggc ccacgctgtc ggcccagccg ccagctcaag cgtcgctttt cgctcgcgtg tgtcaaccgc catctttcgc tcagggcgct cctctgcgtt gcgcattagc caaggagatc ctttgagatt tgagttctcc gtgcaaccgc tgccgtcatc gaacaacaac gacttcgaag tcacagcccg gttcaacaag taccgatcct gatcaagaac cctctcgaag gaaccctgcc ggtcgctggc gaaggagttc gaaaacgctc gcgcatcgag ggagaacgag caccaaggct cgtcgtcgcc ggccgtcgat cgtgtcggct cctgctcgcg gactgagctt gcgcgtcgag 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 197 201038734cacctcaagg aagacgctcc atctcggaca gtcccccgtc cttgaggagt atgctcctcc ctcctcaagg tccaagtttc ggctttgctt ttccagaagg agcaccgccc cagtacaccc aacgatatgg caggtcatgt agcaagaccg ttccgcgacg gccctttacg gccatggcta ggtccgaacg tcgccctcgc ctcaagacgc cacatggaga cccaccggct aagacggccc atgtacgccg ctcgtcaacg tccgccaagg gtcgctctca cctcgcaaga caggagcgcg aacacgaaca gttgtccgcg gagcttcaga aagcacaagg tactccagct cccgtgcagg aaggcggagc atcgagctgg ccgtcgccgg cgcagtccat ccacgctcta gtgcgggcat tcgagcccga acgccgtcaa agtcccgcga aggactcctt ccaagagcat acatcacgac tcatcaacga acatgttcaa agaaggccca ttccccgcaa agtactcgca ccggctttgc ccgctggcct tgcgcgacgc cctcgtccat agaccgtcat agggtttccg acgccgagaa cgtcccccaa tctcccgcca ctggcgcccg agattttccc actcggacat ccgactttga aaacgaccct agcgtatcat cgggcgagct tccaggccct aggctcaggc ccatcctcct tttccaaggg ctgccgcacc aggtcgtgct acatggagct ctgcgctggc caacgtcgag catcaacacc ctccagcttc gcagaccaag cgcgaactcg gaagtgcgtc tasgctcaag cgaggacacg caccagctgg caacaagtcc cgacgtcgcg ggaggaggtg gccctacgcg gacgacgacc cccggctttt catcgaccgc ccccaagaag caagctctcg cactggtgcc cgtggtccac gcagtttcag gattttcagc catgactagc cgttttcatc gggcgacacc ccagctccgc taagtgggag caccctctcc gaacgacggt tgccacccag caccaagtcg cgctatgctg tcaggttgcc ggttcaggtc tgaggtcctc tgagactgag ctcgtcaagc aacccgccca atgaaccgcc ggctttggcg ccctaccgcc ctccagaagc aacacgaaga ggctccgtcc atttccattc gcgctcccga gtggccgccc atgcagtggc atcaacgaca cgcgagagcc gtcgcctcgt gttgcgggcc gaggacctct tccgctgacg gctcccgagg aacagcggcg ctcgcctgcg aaggccctct aacgtcaccg tcggtccagt gagttcggcc agcgtcctca caggcggccg tgagaagctc cttaaggacc gccgctacct cgcactgtca aagaagcagc gccagcgccg tcgaacgctg gaggagctgg tcccctgcga agcgcctccg gcctccaaga cttggtatcg tcgtgatggc acctcgtcga cgtggatcac gcgctaacta tgaacgtttc tctgcgagga acaccgacta ccagccagca ttagcgccat aggagggcgc tgttctcggg cgcagttccg agtcggttaa ccctcgacaa ccgtcggcct actcgctcgg ttaagctcgt gcgccatggc tttgggtcgc tccaggccga acggcgcgtt cggccgtcaa gcggtgtctt ttctcaccca ccaagcaggt ctgttagcgt tgcagatggc cgacccgcat acgttagcaa gctgcgtgc a tccaggacaa accttcagaa cctcggacgc agactggcaa ccgttcgtgt tggactcggg ccggctacga attcgatcaa tctcaagcat tggcactgtc caagccgggc ccacgctgtc ggcccagccg ccagctcaag cgtcgctttt cgctcgcgtg tgtcaaccgc catctttcgc tcagggcgct cctctgcgtt gcgcattagc caaggagatc ctttgagatt tgagttctcc gtgcaaccgc tgccgtcatc gaacaacaac gacttcgaag tcacagcccg gttcaacaag taccgatcct gatcaagaac cctctcgaag gaaccctgcc ggtcgctggc gaaggagttc gaaaacgctc gcgcatcgag ggagaacgag caccaaggct cgtcgtcgcc ggccgtcgat cgtgtcggct cctgctcgcg gactgagctt gcgcgtcgag 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 197 201038734

attctttcgg aggtccaggc ccagctcaac gtggaggcca aggacgttga cgccctgtcg 3480 cgcacccgta cggtcggcga ggtcatcgat gccatgaagg cggagattgc cggcggtcag 3540 cctgctgccc ccgtccaggt cgctgcgccg acgcaggtcg tcgccccggt ccaggcctcc 3600 gcgcctgtcg atagcggcct cctcgccaag gcggagcagg tcgtccttga ggtgctcgct 3660 tccaagactg gttacgagac tgagcttatt gagcttgaca tggagctgga gactgagctt 3720 ggcattgact ccatcaagcg cgtggagatt ctgagcgagg tccaggccca gctcagcgtg 3780 gaggccaagg atgtcgatgc cctctcccgt acgcgcaccg tcggcgaggt cattgacgcg 3840 atgaaggccg agatcgcggg tggtcagccg gccgcccccg tccaggtcgc tgcccctacg 3900 caggtcgtcg ctcccgtcca ggccagcgct cccgtcgact cgggccttct tgctaaggcc 3960 gagcaggtcg tccttgaggt ccttgccagc aagactggct acgagactga gcttattgag 4020 cttgacatgg agcttgagac tgagcttggc atcgactcga ttaagcgcgt cgagatcctc 4080 agcgaggtcc aggcccagct ctccgtcgag gctaaggatg tggatgctct cagccgcacg 4140 cgcacggtgg gcgaggtcat tgatgccatg aaggcggaga tttccggcgg tcagcccgct 4200 gcccccgtcc aggtcgctgc tccgacccag atcgtcgccc cggtccaggt ttcggctccg 4260 gtggacagcg gcctccttgc caaggccgag caggtcgtcc ttgaggtcct cgccagcaag 4320 accggctacg agactgagct gatcgagctt gacatggagc ttgagactga gctgggcatc 4380 gattccatta agcgcgtcga gatcctctcg gaggtccagg cccagctcag cgtggaggcc 4440 aaggatgtcg atgccctctc gcgtacccgt accgtcggcg aggttatcga tgctatgaag 4500 gccgagatca gcggcggtca gcccacggcg cccgttcagg tcgctgcccc tacgcagatc 4560 gttgcccctg tccaggtcag cgctcccgtg gacagcggcc tcctcgctaa ggctgagcag 4620 gtggtgctgg aggtcctggc ctccaagacc ggctacgaga ctgagcttat cgagcttgac 4680 atggagcttg agactgagct tggcattgac agcatcaagc gtgtcgagat cctctccgag 4740 gtgcaggccc agctcagcgt ggaggccaag gacgttgacg cgctcagccg tacgcgcacc 4800 gttggcgagg tgatcgacgc catgaaggcc gagattagcg gtggtcagcc cgctgccccg 4860 gttcaggtgg ctgcccctac gcagatcgtc gcccccgtgc aagcttccgc ccctgtggac 4920 agcggccttc tcgccaaggc cgagcaggtc gtccttgagg tgctggcctc caagaccggc 4980 tacgagactg agctgatcga gcttgacatg gagctggaga ctgagcttgg catcgactcg 5040 atcaagcgcg tggagattct ctcggaggtc caggcccagc tctcggtcga ggccaaggac 5100 gtcgatgcgc tctcccgcac ccgcaccgtg ggcgaggtca tcgacgctat gaaggcggag 5160 atcagcggcg gtcagccggc ggcccctgtg caggtggccg ctccgaccca gatcgtcgct 5220 cctgtccagg tttccgcccc ggtggactcg ggcctcctgg ctaaggccga gcaggtcgtc 5280 cttgaggtcc tcgcttccaa gaccggctac gagactgagc tgatcgagct ggacatggag 5340 cttgagactg agctgggcat cgattcgatc aagcgcgtcg agattctctc ggaggtccag 5400 gcccagctca acgttgaggc caaggacgtg gacgccctct cgcgtactcg caccgttggc 5460 gaggttattg atgctatgaa ggccgagatc gccggtggtc agccggctgc ccctgttcag 5520 gttgctgccc ctgcgccggt ggtcgccccg gtccaggtgt ccaccccggt tgacagcggc 5580 ctccttgcca aggccgagca ggttgtgctg gaggtcctcg cctgcaagac gggctacgag 5640 198 201038734attctttcgg aggtccaggc ccagctcaac gtggaggcca aggacgttga cgccctgtcg 3480 cgcacccgta cggtcggcga ggtcatcgat gccatgaagg cggagattgc cggcggtcag 3540 cctgctgccc ccgtccaggt cgctgcgccg acgcaggtcg tcgccccggt ccaggcctcc 3600 gcgcctgtcg atagcggcct cctcgccaag gcggagcagg tcgtccttga ggtgctcgct 3660 tccaagactg gttacgagac tgagcttatt gagcttgaca tggagctgga gactgagctt 3720 ggcattgact ccatcaagcg cgtggagatt ctgagcgagg tccaggccca gctcagcgtg 3780 gaggccaagg atgtcgatgc cctctcccgt acgcgcaccg tcggcgaggt cattgacgcg 3840 atgaaggccg agatcgcggg tggtcagccg gccgcccccg tccaggtcgc tgcccctacg 3900 caggtcgtcg ctcccgtcca ggccagcgct cccgtcgact cgggccttct tgctaaggcc 3960 gagcaggtcg tccttgaggt ccttgccagc aagactggct acgagactga gcttattgag 4020 cttgacatgg agcttgagac tgagcttggc atcgactcga cgagatcctc 4080 agcgaggtcc aggcccagct ctccgtcgag gctaaggatg tggatgctct cagccgcacg 4140 cgcacggtgg gcgaggtcat tgatgccatg aaggcggaga tttccggcgg tcagcccgct 4200 gcccccgtcc aggtcgctgc tccgacccag atcgtcgccc cggtccaggt ttcggctcc ttaagcgcgt g 4260 gtggacagcg gcctccttgc caaggccgag caggtcgtcc ttgaggtcct cgccagcaag 4320 accggctacg agactgagct gatcgagctt gacatggagc ttgagactga gctgggcatc 4380 gattccatta agcgcgtcga gatcctctcg gaggtccagg cccagctcag cgtggaggcc 4440 aaggatgtcg atgccctctc gcgtacccgt accgtcggcg aggttatcga tgctatgaag 4500 gccgagatca gcggcggtca gcccacggcg cccgttcagg tcgctgcccc tacgcagatc 4560 gttgcccctg tccaggtcag cgctcccgtg gacagcggcc tcctcgctaa ggctgagcag 4620 gtggtgctgg aggtcctggc ctccaagacc ggctacgaga ctgagcttat cgagcttgac 4680 atggagcttg agactgagct tggcattgac agcatcaagc gtgtcgagat cctctccgag 4740 gtgcaggccc agctcagcgt ggaggccaag gacgttgacg cgctcagccg tacgcgcacc 4800 gttggcgagg tgatcgacgc catgaaggcc gagattagcg gtggtcagcc cgctgccccg 4860 gttcaggtgg ctgcccctac gcagatcgtc gcccccgtgc aagcttccgc ccctgtggac 4920 agcggccttc tcgccaaggc cgagcaggtc gtccttgagg tgctggcctc caagaccggc 4980 tacgagactg agctgatcga gcttgacatg gagctggaga ctgagcttgg catcgactcg 5040 atcaagcgcg tggagattct ctcggaggtc caggcccagc tctcggtcga g gccaaggac 5100 gtcgatgcgc tctcccgcac ccgcaccgtg ggcgaggtca tcgacgctat gaaggcggag 5160 atcagcggcg gtcagccggc ggcccctgtg caggtggccg ctccgaccca gatcgtcgct 5220 cctgtccagg tttccgcccc ggtggactcg ggcctcctgg ctaaggccga gcaggtcgtc 5280 cttgaggtcc tcgcttccaa gaccggctac gagactgagc tgatcgagct ggacatggag 5340 cttgagactg agctgggcat cgattcgatc aagcgcgtcg agattctctc ggaggtccag 5400 gcccagctca acgttgaggc caaggacgtg gacgccctct cgcgtactcg caccgttggc 5460 gaggttattg atgctatgaa ggccgagatc gccggtggtc agccggctgc ccctgttcag 5520 gttgctgccc ctgcgccggt ggtcgccccg gtccaggtgt ccaccccggt tgacagcggc 5580 ctccttgcca aggccgagca ggttgtgctg gaggtcctcg cctgcaagac gggctacgag 5640 198 201038734

actgagctta tcgagcttga catggagctg gagactgagc ttggcatcga ctccatcaaa 5700 cgcgtcgaga ttctttcgga ggtccaggcc cagctgtcgg tggaggctaa ggatgtcgat 5760 gccctcagcc gcacgcgcac ggtcggtgag gtcatcgatg ctatgaaggc cgagatttcg 5820 ggcggtcagc ccaccgcccc cgtgcaggtc gccgcgccca cccaggtcgt ggccccggtc 5880 aaggtttcca cgcccgtgga ctcgggcctt ctcgccaagg ccgagcaggt cgtgctggag 5940 gttctcgcct ccaagacggg ttacgagact gagctgattg agcttgacat ggagctggag 6000 actgagctgg gcattgactc catcaagcgc gtcgagatcc tctcggaggt ccaggcccag 6060 ctcaacgtcg aggccaagga cgtcgatgcc ctctcgcgca cccgcaccgt cggcgaggtc 6120 attgatgcca tgaaggccga gatcgctggc gatcagcctg ccccggctgt ggtcccggtg 6180 caggccaagt cgggtgtcgc gaaccccgcc ctcctcgcca aggcggagca ggtcgtgctg 6240 gaggtcctgg ccagcaagac gggctacgag actgagctta tcgagcttga catggagctt 6300 gagactgagc ttggtattga ctcgattaag cgcgttgaga tcctttccga ggtccaggcc 6360 gagctgtccg tggaggccaa ggatgtcgat gcgctctccc gcacccgcac ggtgggcgag 6420 gtcatcgacg ctatgaaggc cgagattgcc ggctccgcgg tcactgtcgc tacccttgac 6480 gactcgacca ttatggagga gactgacgac gaggacgagg actttatcct gtacgaccac 6540 gtctacggct ccgagtgcga ggatctctcg ctctcgttct cgtcggtcaa gtcgattcct 6600 cgcgcggaca agctcctgct ggacaacatt gccgagcgcc ccattgtcat tgtcgattgc 6660 ggcacgaagc tcacgaccga gctggcgaag gccatcggcg agcgcgctgt cgttgccacg 6720 ttctcggccc agtcgctcgt gtcccgtggc ttcgtgggca agagcttcac cctcggcaac 6780 accgaggagt cggagatcga gaagatggtg tcctccatcg agtcgtccta cggcaagatc 6840 ggcggctttg tctaccagca ctttcatgac agcgactacg gtatgcagct cggctgggct 6900 ctcatggccg cgaagcacct caaggagtcc ctcaacgacc cgatcaagaa cggccgcacc 6960 tttttcctgg ctgtcgcccg catgaacggc aagctcggca tggacaacgc ctccgtccac 7020 gaccagggca tcgtcgagag ctgcggtatc gctgagcgtg gtgccatctt tggcctctgc 7080 aagaccctgg acctggagtg gcctaacgtg tttgcgcgcg gtgtggacat cgcggagggc 7140 atgtcctact ccctcgcggc cgagctgatc gtcgatgaga tcagctgcgc caacctttcg 7200 atccgcgaga gcggctacac tattagcggc gagcgcttca ccacggaggc gcacaagctc 7260 gtcacgggca agcctcacgc gcccatcaag aagaaggacg cctttctcgt gtcgggtggt 7320 gctcgcggca tcacgcccct gtgcattcgc gagattgcca aggccgtcaa gggtggcacc 7380 tacattctca tgggccgctc ggcgctcgcg gacgagcccc tctgggctaa cggcaagagc 7440 ggcaaggacc tcgacaaggc cggcctcgcc ttccttaagg aggagttcgc tgccggccgt 7500 ggctcgaagc ccacccccaa ggtccacaag tcgctcatcg acaaggtcct cggcatccgc 7560 gaggttcgcg cgtccatcgc caacatcgag gcgcacggcg ctaaggccat ctacctctcg 7 620 tgcgatgtgt cgagcgccga gaaggtcaag gctgccgtcc agaaggtcga gaaggagcat 7 680 ctcgtccgca tcacgggcat cgtgcacgcc tccggcgtcc tgcgcgacaa gctcgtcgag 7740 aacaagaccc tcgacgactt taacgctgtg tacggcacga aggtcacggg cctcgtcaac 7 8〇0 ctccttagcg ccgtcaacat gaacttcgtc cgccacctgg tgatgttctc gtcgctcgct 7860 ggttaccacg gcaacgtcgg ccagtcggac tacgctatgg ccaacgagag ccttaacaag 7920 199 201038734 atcggcttcc ccttgggatg cagatcatcc cccagccagg gccaccatcg cacggcaaga aacttttacg gtcatcgacc ggcaaggtca gcctacaagg ttttcgctcc ttccacggcc acgacccagt gagccggtca aacctccgta atcgcgcccg gttctcaagt gccagcgtcg gtcttggtgc gcggcatggt ctcgcgaggg tcctcgtcgg tccagacctt acgtcctgcc ccggccacca acgccgtgca aggtccaggc ccgtcatcgt aggagtccaa cgctgttccg gcaccaacat accctttcat actccgccag gcgagaagta ccgttttcta tggtcaacga cgcgtactcc gacgccggcg tggcgccgag caactggggc tacccctgag catgacggtc cctctggggc ggcccaggtc ggtgctcacc gctcggcaag ctcgcgctcg tggcatcacc cgatctcacc ggcggacgcc ccttccgaac ctacaccacg catgcatgac taagatggaa cagctctgcg ctcaagaagc actgtcgctc gtcccgcccg cttaacccct gccattggtt gtggaggacg aagctcactg gcctcgaacg acttcccgtc gccgacgagc aagctcctca gccactgagc gccttccagg aactgcgagc ctgcaggccc gagcagggcg ttctaa tcaagtccat agttccagtc gcattgtgct tcagccccct tcctcaagtc acctcgccca cgcagctctt agcagagcct acaacggcaa cggccttcat tgtacgacgg acgtgtccga gcggccagtt ccatgctcgt gcgtcgatat tcggcaacac aggtgttcct ctgcttcggc catgggcgtt ctcgtccaac ctccaagtcg ccaccagatc cctcgtgaag ctccggcgtc ggatgacgat gatggtgccg cctcaaggac caagaccctg cactagcctc tgccgacatc ctgggtccgc ctacaagccc ctccggctcg ctcgggccgc 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 8700 87 60 8820 8880 8940 8976actgagctta tcgagcttga catggagctg gagactgagc ttggcatcga ctccatcaaa 5700 cgcgtcgaga ttctttcgga ggtccaggcc cagctgtcgg tggaggctaa ggatgtcgat 5760 gccctcagcc gcacgcgcac ggtcggtgag gtcatcgatg ctatgaaggc cgagatttcg 5820 ggcggtcagc ccaccgcccc cgtgcaggtc gccgcgccca cccaggtcgt ggccccggtc 5880 aaggtttcca cgcccgtgga ctcgggcctt ctcgccaagg ccgagcaggt cgtgctggag 5940 gttctcgcct ccaagacggg ttacgagact gagctgattg agcttgacat ggagctggag 6000 actgagctgg gcattgactc catcaagcgc gtcgagatcc tctcggaggt ccaggcccag 6060 ctcaacgtcg aggccaagga cgtcgatgcc ctctcgcgca cccgcaccgt cggcgaggtc 6120 attgatgcca tgaaggccga gatcgctggc gatcagcctg ccccggctgt ggtcccggtg 6180 caggccaagt cgggtgtcgc gaaccccgcc ctcctcgcca aggcggagca ggtcgtgctg 6240 gaggtcctgg ccagcaagac gggctacgag actgagctta tcgagcttga catggagctt 6300 gagactgagc ttggtattga ctcgattaag cgcgttgaga tcctttccga ggtccaggcc 6360 gagctgtccg tggaggccaa ggatgtcgat gcgctctccc gcacccgcac ggtgggcgag 6420 gtcatcgacg ctatgaaggc cgagattgcc ggctccgcgg tcactgtcgc tacccttga c 6480 gactcgacca ttatggagga gactgacgac gaggacgagg actttatcct gtacgaccac 6540 gtctacggct ccgagtgcga ggatctctcg ctctcgttct cgtcggtcaa gtcgattcct 6600 cgcgcggaca agctcctgct ggacaacatt gccgagcgcc ccattgtcat tgtcgattgc 6660 ggcacgaagc tcacgaccga gctggcgaag gccatcggcg agcgcgctgt cgttgccacg 6720 ttctcggccc agtcgctcgt gtcccgtggc ttcgtgggca agagcttcac cctcggcaac 6780 accgaggagt cggagatcga gaagatggtg tcctccatcg agtcgtccta cggcaagatc 6840 ggcggctttg tctaccagca ctttcatgac agcgactacg gtatgcagct cggctgggct 6900 ctcatggccg cgaagcacct caaggagtcc ctcaacgacc cgatcaagaa cggccgcacc 6960 tttttcctgg ctgtcgcccg catgaacggc aagctcggca tggacaacgc ctccgtccac 7020 gaccagggca tcgtcgagag ctgcggtatc gctgagcgtg gtgccatctt tggcctctgc 7080 aagaccctgg acctggagtg gcctaacgtg tttgcgcgcg gtgtggacat cgcggagggc 7140 atgtcctact ccctcgcggc cgagctgatc gtcgatgaga tcagctgcgc caacctttcg 7200 atccgcgaga gcggctacac tattagcggc gagcgcttca ccacggaggc gcacaagctc 7260 gtcacgggca agcctcacgc gcccatcaag aagaaggacg cctttctcgt g tcgggtggt 7320 gctcgcggca tcacgcccct gtgcattcgc gagattgcca aggccgtcaa gggtggcacc 7380 tacattctca tgggccgctc ggcgctcgcg gacgagcccc tctgggctaa cggcaagagc 7440 ggcaaggacc tcgacaaggc cggcctcgcc ttccttaagg aggagttcgc tgccggccgt 7500 ggctcgaagc ccacccccaa ggtccacaag tcgctcatcg acaaggtcct cggcatccgc 7560 gaggttcgcg cgtccatcgc caacatcgag gcgcacggcg ctaaggccat ctacctctcg 7 620 tgcgatgtgt cgagcgccga gaaggtcaag gctgccgtcc agaaggtcga gaaggagcat 7 680 ctcgtccgca tcacgggcat cgtgcacgcc tccggcgtcc tgcgcgacaa gctcgtcgag 7740 aacaagaccc tcgacgactt taacgctgtg tacggcacga aggtcacggg cctcgtcaac 7 8〇0 ctccttagcg ccgtcaacat gaacttcgtc cgccacctgg tgatgttctc gtcgctcgct 7860 ggttaccacg gcaacgtcgg ccagtcggac tacgctatgg ccaacgagag ccttaacaag 7920 199 201038734 atcggcttcc ccttgggatg cagatcatcc cccagccagg gccaccatcg cacggcaaga aacttttacg gtcatcgacc ggcaaggtca gcctacaagg ttttcgctcc ttccacggcc acgacccagt gagccggtca aacctccgta atcgcgcccg gttctcaagt gccagcgtcg gtcttggtgc gcggcatggt ctcgcgaggg Tcctcgtcgg tcca gacctt acgtcctgcc ccggccacca acgccgtgca aggtccaggc ccgtcatcgt aggagtccaa cgctgttccg gcaccaacat accctttcat actccgccag gcgagaagta ccgttttcta tggtcaacga cgcgtactcc gacgccggcg tggcgccgag caactggggc tacccctgag catgacggtc cctctggggc ggcccaggtc ggtgctcacc gctcggcaag ctcgcgctcg tggcatcacc cgatctcacc ggcggacgcc ccttccgaac ctacaccacg catgcatgac taagatggaa cagctctgcg ctcaagaagc actgtcgctc gtcccgcccg cttaacccct gccattggtt gtggaggacg aagctcactg gcctcgaacg acttcccgtc gccgacgagc aagctcctca gccactgagc gccttccagg aactgcgagc ctgcaggccc gagcagggcg ttctaa tcaagtccat agttccagtc gcattgtgct tcagccccct tcctcaagtc acctcgccca cgcagctctt agcagagcct acaacggcaa cggccttcat tgtacgacgg acgtgtccga gcggccagtt ccatgctcgt gcgtcgatat tcggcaacac aggtgttcct ctgcttcggc catgggcgtt ctcgtccaac ctccaagtcg ccaccagatc cctcgtgaag ctccggcgtc ggatgacgat gatggtgccg cctcaaggac caagaccctg cactagcctc tgccgacatc ctgggtccgc ctacaagccc ctccggctcg ctcgggccgc 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 8700 87 60 8820 8880 894 0 8976

&lt;210〉 121 &lt;211〉 5880 &lt;212&gt; DNA &lt;213&gt; 人工序列 &lt;220&gt; &lt;223&gt; 密碼子最佳化的ΡΕΆ2 &lt;400&gt; 121 atggtgaagc tttccgttgg tgacaacatt tgccacgatc agcgcgtcgc cgtggtcggc 60 atggccgtca tgtacgccgg ctgccagaac cagcacgagt tttggcagag cctccagggt 120&lt;210> 121 &lt;211> 5880 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Codon-optimized ΡΕΆ2 &lt;400&gt; 121 atggtgaagc tttccgttgg tgacaacatt tgccacgatc agcgcgtcgc cgtggtcggc 60 atggccgtca tgtacgccgg Ctgccagaac cagcacgagt tttggcagag cctccagggt 120

aagaacatga acagcaagag catcagccag aaccgcctgg gctccgagta ccgcgaggag 180 cactttaagc cggagcgctc gaagtacagc gacaccttct gcaacgagcg ttacggctgc 240 atcgacgaga acgtccagag cgagcatgag ctcctcctga agctcgctaa ggacgcgatc 300 gccgatacca agggcagcat cgaccttaac aagaccggca ttgtctccgg ctgcctctcg 360 ttccctatgg ataacctcca gggcgacctt ctcaacctct accagtgcca tattgagaag 420 aagatcggcc cgaacgccct caaggatgtc aacctctggt cgaagcgcac gaccaacggt 480 aaggacgata agaaggccta cttcgatccc gccagcttcg tcgctgagca gcttgacatg 540 ggtcccctcc actactcgct cgacgctgcc tgcgcctccg ctctctacgt cctccgcctc 600 gcccaggacc acctcctcag cggtgccgcc gacaccatgc tctgcggcgc ctcgtgcctc 660 ccggagccct ttttcatcct ttcgggcttt tcgaccttcc acgccatgcc cctttcgggt 720 gacgtgtcgg cccctcttca caagacgagc cagggcctca ctccgggcga gggcggtgct 780 atcatggtcc tgaagcgcct caacgatgcc attcgcgacg gcgaccgcat ctacggcacg 840 200 201038734aagaacatga acagcaagag catcagccag aaccgcctgg gctccgagta ccgcgaggag 180 cactttaagc cggagcgctc gaagtacagc gacaccttct gcaacgagcg ttacggctgc 240 atcgacgaga acgtccagag cgagcatgag ctcctcctga agctcgctaa ggacgcgatc 300 gccgatacca agggcagcat cgaccttaac aagaccggca ttgtctccgg ctgcctctcg 360 ttccctatgg ataacctcca gggcgacctt ctcaacctct accagtgcca tattgagaag 420 aagatcggcc cgaacgccct caaggatgtc aacctctggt cgaagcgcac gaccaacggt 480 aaggacgata agaaggccta cttcgatccc gccagcttcg tcgctgagca gcttgacatg 540 ggtcccctcc actactcgct cgacgctgcc tgcgcctccg ctctctacgt cctccgcctc 600 gcccaggacc acctcctcag cggtgccgcc gacaccatgc tctgcggcgc ctcgtgcctc 660 ccggagccct ttttcatcct ttcgggcttt tcgaccttcc acgccatgcc cctttcgggt 720 gacgtgtcgg cccctcttca caagacgagc cagggcctca ctccgggcga gggcggtgct 780 atcatggtcc tgaagcgcct caacgatgcc attcgcgacg gcgaccgcat ctacggcacg 840 200 201038734

❹ ctcctgggcg ccgagctttc caacgcgggt tgcggcctcc cgctctcccc gcacatgccg 900 tccgagttcg actgcatgga gaaggccctc cagcgcgttc accgcctccc gtcctccatc 960 cagtacgtgg agtgccacgc cactggcacc ccgcagggcg acaaggtcga gatcgacgcc 1020 atgacgaagt gcttcggcga gcatctgcct cgcttcggct ccaccaaggg taacttcggc 1080 cacaccctcg tggctgctgg ctttgcgggc atgtgcaagg tcctcctctc gatgcagtac 1140 ggtgagattc ctcctacgcc tggcctggag aaccccgaca acattatgca cgatcttgtc 1200 gttaccgaga ctattccctg gccgaacacc aacggcgatc ttaagcgtgc gtgcctcagc 1260 gcctttggct ttggcggtac taacgcccac gccgtgttcg aggagtaccg cagcgacctt 1320 caggccaaca agacccttga gaacgagagc aagtcccacg agatcttttc ctcctttaag 1380 attgccattg ttggcatgga gtccgagttt ggcactctca agggcctcca ggagttcgag 1440 cgtgccatct acaacggcgg ccacggcgcg tgcgaccttc cggagaaccg ctggcgcttt 1500 ctcggtgagg acaaggagtt tctccaggcc tgcggcctcc agaagctccc gcgtggctgc 1560 tacatcaagg aggtcgagac tgactttaag cgccttcgcc tccccatgat ccaggaggac 1620 atcctccgcc ccctccagct cctcgccgtg tcgatcatcg accgcgccct caacgccagc 1680 ggcgttaagc ccaacggcaa ggtcgccgtc ctcgtgggcc tcggcaccga tcttgagctc 1740 taccgccacc gcgctcgcgt cgccctgaag gagcgccttc agaccgccgt caaggaggac 1800 atccccctgc tggagaagct catgaactac gtgaacgacc gcggcacctc cacgtcctac 1860 acctcgtaca tcggcaacct cgttgcgacc cgcgtcagct cgctctgggg cttcaccggc 1920 cctagcttca cgatcacgga gggcgagaac tcggtttacc gttgcctcga cctcggccgc 1980 tggttcctcg ccaacggtga ggtcgatgcc gtggttgtcg ctggcgtgga tctctgcggc 2040 tcggccgaga acctgttcgt caagtcgcgc cgctccaagg tgtccaccca gaacgagccc 2100 tttgctaact ttgagtcgaa cgccgacggc tacttcgccg gcgacggctg cggtgccctc 2160 gttctcaagc gcctttcgga ctgcactgac tccaccgaga agatctacgc gaccgtggac 2220 agcattgctg tcggcgacga ggtgggcccg actattaagc aggccctgaa gaacgcctcg 2280 atcgccgcga aggacatcga gctcgcggag ctctccgcct ccagcggcaa gcaccactcc 2340 ggccgcatca cctgcgagga cgagcttaac gagctcggcg agatcttcaa cgagggcatt 2400 cagcgcgtgg ccatcggcag cgtcaaggcc aacgtcggcg acgtcggcta cgcctccggt 2460 gctgccagcc tcatcaagac ggccctctgc ctctacaacc gctacctccc caagctcccc 2520 aactggaaca agccgaccaa ggacgtcgag tggtcgaaga gcttctttgt ctgcgagcac 2580 tcgcgcgcct ggctcaagaa cgtggacgag aaccgccacg cggtcgtgag cggcgtctgc 2640 gagaacggct cctgctacgg catcgtcatg agcgacgtcc agggccacca tgaggagtcg 2700 aacctcgtgt ccctcgataa gaacgagccc aaggtgctcg gtatctacgg cgattccgtg 2760 gacgatattc tggtccagct gaacaagtac ctggagaagt tccttcagga gactggcact 2820 gctgcggctg cgcagaaggt gaagagccct accattgaca tcgactcgaa cgtctttgcc 2880 gagatgctga accttcccca ggacaagaac aagaagtttg ccgtcgctct ggtcacgacc 2940 cccaacaagc tccagcgcga gattgagctc gccgttaagg gcatccctcg ctgcgtgaag 3000 gccaagcgcg actggtgctc cccctccggc agcatctttg cgtgcaaccc gctcaagtcg 3060 gacaacattg cctttatgta cggcgagggc cgctcgcctt acgccggcct cggctacgat 3120 201 201038734 ctccaccgca tctggcccat gcttcacgag ctcgtgaaca accgcacgac tgagctgtgg 3180 gaccagggtg actcgtggta cctgccgcgc agctcctccg tggccgagaa ggagaaggtc 3240 tttggcgact tcgacaagaa ccagatcgag atgttccgcc tcggtatttt cgtcagcatg 3300 tgctttaccg acatggcgac ggagctcctc ggccttaagc cgaaggccgc tttcggcctc 3360 tccctcggcg agatcagcat gctctttgct ttctcgaaga agaacaccaa gctctccaag 3420 gagcttactc gccgcctcaa ggaggccaag gtgtgggcgt cgcagctggc cgtcgagttc 3480 gccgccatcc gcgacctttg gaacatcccg gccgacaagt ccatcgatga gttctggcag 3540 ggttacttcg tttacgccaa ccgtacgctc gtggags^cs ccattggcga gaacaagttc 3600 gtccgcctcc ttatcgtcaa cgactcccag tcctgcctca ttgccggtaa gcccgatgag 3660 tgccagaagg tcatcgagaa gctccacctt aagctccccg ccgtccccgt cacccagggc 3720 atgattggcc actgcccgga ggccattccc tacctcgacc agatcagcca catccacgag 3780 atgcttgaga tcccgaagcc tgagaacgtc aagctcttca cgacgtccga gaaccgcgag 3840 cttgtctcga tgaaggactc cgttagcaag ctcgtcgcgg agatctacca gcacgtcgct 3900 gacttcccca acattgtcaa caaggtcaag gagacttgca agacggacat tttcatcgag 3960 ctgggcagca acaactaccg ttccggtgcc gtcaagacta tcctcggtcc ggagatcgtg 4020 agcgttgcca tcgaccgtca gaacgagact gcctggggcc agctcatgaa gatggtcgcc 4080 agcctgatct cccaccgcgt ccccggcgtc gagctcaaga agctgtacca tccggagctc 4140 ctgaagttcg atccccaggc caagcccaac cgctttatcc gcaacatcga gctcaacggc 4200 tttttcgacc gcacgaacat catcgtcgat aagcagcttt cccctgcgga cccgaagctc 4260 gccgagatcg tcaacaaccg caacatgccg aaggataacg tgtacgtccc cattgagcgc 4320 gtcaagacga tgatcaaggc cgagcccgct aacctccagg tgtccgtcgg ctcgaagccc 4380 gtggtcaccg agcgtatctc gtcggacgac aacctctttg agaagctctc ggagatcact 4440 aagtccttcg acggtgtcaa cgcctgcacc gaggccatgc tcggcgattc gggctttctc 4500 aagacgtacg aggttgacta cccgctctac accggcgcta tggccaaggg tatcgcctcc 4560 gccgacctcg tcattgcggc gggtaagtcg aagatccttg cgtcctttgg tgctggcggc 4620 ctcgctctcc aggtggtcga ggatgccatt aagcagatca aggctgagct tggcaacggt 4680 ccctttgccg tcaacctcat ccactcgcct ttcgacccct cgcttgagaa gggcaacgtt 4740 gaccttttcc tcaagtacaa cgtccgcttt gtcgaggtga gcgcgttcat gagcctcacc 4800 ccccaggtcg ttcgctaccg cgctgccggc cttgccaagg cccgtgacgg ctcggtcaag 4860 attcagaacc gcatcatcgc caagatttcg cgcacggagc tggccgagct cttcctcaag 4920 cccgctccga agaacatcct cgatgccctc gttgccgacg gctcgatttc ccaggagcag 4980 gctcagctcg cgctcctcgt ccctatggcc gatgacatca ccgttgaggc cgactccggt 5040 ggccacaccg acaaccgccc cattcatgtg ctcctccccc tcatcatcca gcagcgcaac 5100 cgcatttgca agcagtaccc gaagcacctc aaggtccgca tcggcgctgc cggtggcatc 5160 ggttgcccta aggcggcttt tgccgccttt gagatgggtg cggcctacat cgccacgggc 5220 accgttaacc agctctcgaa ggaggccggc acctgcgact acgtgcgcaa ggtgctcaac 5280 aaggccacct actccgacgt cacgatggct cccgctgccg acatgttcga ccacggtgtc 5340❹ ctcctgggcg ccgagctttc caacgcgggt tgcggcctcc cgctctcccc gcacatgccg 900 tccgagttcg actgcatgga gaaggccctc cagcgcgttc accgcctccc gtcctccatc 960 cagtacgtgg agtgccacgc cactggcacc ccgcagggcg acaaggtcga gatcgacgcc 1020 atgacgaagt gcttcggcga gcatctgcct cgcttcggct ccaccaaggg taacttcggc 1080 cacaccctcg tggctgctgg ctttgcgggc atgtgcaagg tcctcctctc gatgcagtac 1140 ggtgagattc ctcctacgcc tggcctggag aaccccgaca acattatgca cgatcttgtc 1200 gttaccgaga ctattccctg gccgaacacc aacggcgatc ttaagcgtgc gtgcctcagc 1260 gcctttggct ttggcggtac taacgcccac gccgtgttcg aggagtaccg cagcgacctt 1320 caggccaaca agacccttga gaacgagagc aagtcccacg agatcttttc ctcctttaag 1380 attgccattg ttggcatgga gtccgagttt ggcactctca agggcctcca ggagttcgag 1440 cgtgccatct acaacggcgg ccacggcgcg tgcgaccttc cggagaaccg ctggcgcttt 1500 ctcggtgagg acaaggagtt tctccaggcc tgcggcctcc agaagctccc gcgtggctgc 1560 tacatcaagg aggtcgagac tgactttaag cgccttcgcc tccccatgat ccaggaggac 1620 atcctccgcc ccctccagct cctcgccgtg tcgatcatcg accgcgccct caacgcc agc 1680 ggcgttaagc ccaacggcaa ggtcgccgtc ctcgtgggcc tcggcaccga tcttgagctc 1740 taccgccacc gcgctcgcgt cgccctgaag gagcgccttc agaccgccgt caaggaggac 1800 atccccctgc tggagaagct catgaactac gtgaacgacc gcggcacctc cacgtcctac 1860 acctcgtaca tcggcaacct cgttgcgacc cgcgtcagct cgctctgggg cttcaccggc 1920 cctagcttca cgatcacgga gggcgagaac tcggtttacc gttgcctcga cctcggccgc 1980 tggttcctcg ccaacggtga ggtcgatgcc gtggttgtcg ctggcgtgga tctctgcggc 2040 tcggccgaga acctgttcgt caagtcgcgc cgctccaagg tgtccaccca gaacgagccc 2100 tttgctaact ttgagtcgaa cgccgacggc tacttcgccg gcgacggctg cggtgccctc 2160 gttctcaagc gcctttcgga ctgcactgac tccaccgaga agatctacgc gaccgtggac 2220 agcattgctg tcggcgacga ggtgggcccg actattaagc aggccctgaa gaacgcctcg 2280 atcgccgcga aggacatcga gctcgcggag ctctccgcct ccagcggcaa gcaccactcc 2340 ggccgcatca cctgcgagga cgagcttaac gagctcggcg agatcttcaa cgagggcatt 2400 cagcgcgtgg ccatcggcag cgtcaaggcc aacgtcggcg acgtcggcta cgcctccggt 2460 gctgccagcc tcatcaagac ggccctctgc ctctacaacc gctacctccc caagctcccc 2520 aactggaaca agccgaccaa ggacgtcgag tggtcgaaga gcttctttgt ctgcgagcac 2580 tcgcgcgcct ggctcaagaa cgtggacgag aaccgccacg cggtcgtgag cggcgtctgc 2640 gagaacggct cctgctacgg catcgtcatg agcgacgtcc agggccacca tgaggagtcg 2700 aacctcgtgt ccctcgataa gaacgagccc aaggtgctcg gtatctacgg cgattccgtg 2760 gacgatattc tggtccagct gaacaagtac ctggagaagt tccttcagga gactggcact 2820 gctgcggctg cgcagaaggt gaagagccct accattgaca tcgactcgaa cgtctttgcc 2880 gagatgctga accttcccca ggacaagaac aagaagtttg ccgtcgctct ggtcacgacc 2940 cccaacaagc tccagcgcga gattgagctc gccgttaagg gcatccctcg ctgcgtgaag 3000 gccaagcgcg actggtgctc cccctccggc agcatctttg cgtgcaaccc gctcaagtcg 3060 gacaacattg cctttatgta cggcgagggc cgctcgcctt acgccggcct cggctacgat 3120 201 201038734 ctccaccgca tctggcccat gcttcacgag ctcgtgaaca accgcacgac tgagctgtgg 3180 gaccagggtg actcgtggta cctgccgcgc agctcctccg tggccgagaa ggagaaggtc 3240 tttggcgact tcgacaagaa ccagatcgag atgttccgcc tcggtatttt cgtcagcatg 3300 tgctttaccg acatggcgac ggagctcctc ggccttaagc cgaaggccgc tttcggcctc 3360 tccctcggcg agatcagcat gctctttgct ttctcgaaga agaacaccaa gctctccaag 3420 gagcttactc gccgcctcaa ggaggccaag gtgtgggcgt cgcagctggc cgtcgagttc 3480 gccgccatcc gcgacctttg gaacatcccg gccgacaagt ccatcgatga gttctggcag 3540 ggttacttcg tttacgccaa ccgtacgctc gtggags ^ cs ccattggcga gaacaagttc 3600 gtccgcctcc ttatcgtcaa cgactcccag tcctgcctca ttgccggtaa gcccgatgag 3660 tgccagaagg tcatcgagaa gctccacctt aagctccccg ccgtccccgt cacccagggc 3720 atgattggcc actgcccgga ggccattccc tacctcgacc agatcagcca catccacgag 3780 atgcttgaga tcccgaagcc tgagaacgtc aagctcttca cgacgtccga gaaccgcgag 3840 cttgtctcga tgaaggactc cgttagcaag ctcgtcgcgg agatctacca gcacgtcgct 3900 gacttcccca acattgtcaa caaggtcaag gagacttgca agacggacat tttcatcgag 3960 ctgggcagca acaactaccg ttccggtgcc gtcaagacta tcctcggtcc ggagatcgtg 4020 agcgttgcca tcgaccgtca gaacgagact gcctggggcc agctcatgaa gatggtcgcc 4080 agcctgatct cccaccgcgt ccccggcgtc gagctcaaga agctgtacca tccggagctc 4140 ctgaagttcg atccccaggc caa gcccaac cgctttatcc gcaacatcga gctcaacggc 4200 tttttcgacc gcacgaacat catcgtcgat aagcagcttt cccctgcgga cccgaagctc 4260 gccgagatcg tcaacaaccg caacatgccg aaggataacg tgtacgtccc cattgagcgc 4320 gtcaagacga tgatcaaggc cgagcccgct aacctccagg tgtccgtcgg ctcgaagccc 4380 gtggtcaccg agcgtatctc gtcggacgac aacctctttg agaagctctc ggagatcact 4440 aagtccttcg acggtgtcaa cgcctgcacc gaggccatgc tcggcgattc gggctttctc 4500 aagacgtacg aggttgacta cccgctctac accggcgcta tggccaaggg tatcgcctcc 4560 gccgacctcg tcattgcggc gggtaagtcg aagatccttg cgtcctttgg tgctggcggc 4620 ctcgctctcc aggtggtcga ggatgccatt aagcagatca aggctgagct tggcaacggt 4680 ccctttgccg tcaacctcat ccactcgcct ttcgacccct cgcttgagaa gggcaacgtt 4740 gaccttttcc tcaagtacaa cgtccgcttt gtcgaggtga gcgcgttcat gagcctcacc 4800 ccccaggtcg ttcgctaccg cgctgccggc cttgccaagg cccgtgacgg ctcggtcaag 4860 attcagaacc gcatcatcgc caagatttcg cgcacggagc tggccgagct cttcctcaag 4920 cccgctccga agaacatcct cgatgccctc gttgccgacg gctcgatttc ccaggagcag 4980 gctcagctcg cgctcc tcgt ccctatggcc gatgacatca ccgttgaggc cgactccggt 5040 ggccacaccg acaaccgccc cattcatgtg ctcctccccc tcatcatcca gcagcgcaac 5100 cgcatttgca agcagtaccc gaagcacctc aaggtccgca tcggcgctgc cggtggcatc 5160 ggttgcccta aggcggcttt tgccgccttt gagatgggtg cggcctacat cgccacgggc 5220 accgttaacc agctctcgaa ggaggccggc acctgcgact acgtgcgcaa ggtgctcaac 5280 aaggccacct actccgacgt cacgatggct cccgctgccg acatgttcga ccacggtgtc 5340

202 201038734202 201038734

gagctccagg ttctcaagaa gggcaccatg tttccgtcgc gcgccaagaa gctctacgac 5400 ctctttaaga agtacaagtc gatcgaggag ctccctgccg acgaggtcaa gaagctggag 5460 cagaaggttt ttaagaagtc gttcgacgag gtctgggacg agactaagaa ctactacatt 5520 aaccgcctcc actcccctga gaagatcgag cgcgcggagc gtgacgccaa gctgaagatg 5580 tcgctctgct ttcgttggta cctgagcaag tcgtcccgct gggccaacac cggcgagtcg 5640 ggccgtgtcc aggactacca gatctggtgc ggccccgcca tcggctcgta caacgacttc 5700 gcgaagggct cgccctgcct tgaccctgag atccttggct cgttcccgtc ggttgtccag 5760 atcaacaagc atattctgcg cggcgcttgc ttctaccagc gtctttcgca gctcaagtac 5820 cttaacttca actacgagga gctcgatacg ctcacctaca gcgctagcaa ctttatctaa 5880 &lt;210&gt; 122 &lt;211&gt; 4368 &lt;212&gt; DNA &lt;213&gt; 人工序列 &lt;220&gt; &lt;223&gt; 密碼子最佳化的PFA3 &lt;400&gt; 122 atggttggcc tgcagatgaa gaagaagcct gtgtgggaga tgtcgaagga ggagcagtcg 60 tccggcaaga acgtcgtctt tgactacgac gagctcctcg aattcgcgga gggtgacatc 120 ggcaaggtgt tcggccccaa gtttgacatc atcgacaagt acagccgccg tgtgcgcctc 180 ccggcccgcg agtacctcct cgtcacccgt gtcacgctca tggatgccga ggtcggcaac 240 ttccgcgtcg gctcgcgcat ggtcaccgag tacgacgtcc cggtgaacgg cgagctttcc 300 cagggcggcg acgttccctg ggccgtcctc gtcgagtcgg gccagtgcga cctcatgctt 360 atctcgtaca tgggcattga ctttcagtgc aagggtgacc gcgtttaccg ccttctcaac 420 acgaccctca cgttctacgg tgtcgcccac gagggcgaga ctctcgttta cgacatccgc 480 gtcactggtt tcgccaaggg catgcacggc gagattagca tgttcttctt cgagtacgac 540 tgctacgtca acggccgcct gctcatcgag atgcgcgacg gttgcgctgg cttcttcacg 600 gacgaggagc tcgccgcggg caagggcgtc atcaagaccg tcgctgagct ccacaagcgc 660 aagtcgattg tgcccaagtc gatcaagcct tttgccctca accccgccgt ccacaagacg 720 atgttcagcg agaacgacat ggagaagctt tgcgagcgcc agtgggagaa cgtcctcggc 780 tccggcctcc agggcatcga ctacaagctg tgcgcccgca agatgctcat gatcgaccgc 840 atcacgaaga tccagcacaa cggcggtgcg tacggcctcg gcctcctcgt tggcgagaag 900 attcttgagc gcgaccattg gtacttccct tgccacttcg tcaaggacca ggtgatggcg 960 ggctccctcg ttagcgacgg ctgctcgcag ctgctcaagc tttacatgct ttggctcggc 1020 ctccacgacg tggtccccga tttccagttc cgtcctgtcc ctggccagcc caacaaggtg 1080 cgctgccgtg gccagatcag cccccatcgt ggcaagctcg tgtacgtgat ggagattcgc 1140 gagatgggtt tcaacgagtc caccggccag ccctacgcga tcgctgacgt tgacattatc 1200 gatgtgaact acgagctcgg ccagtccttt gacatggccg acatcgactc gtacggccgt 1260 ggcaacctct ccaagaagat tgtcgtcgat ttcaagggca ttgcgctcca gatggagggc 1320 accgtcaaga gctccaacat catcgattcg tcccccaagt ccacgattat ccagccgccg 1380 cccaactgcc tccgcggcga tcctctcgcc ccctcgcagg tcacctggca cccgatggcc 1440 203 201038734 ggtgtcaacg gcgcccccgc cccctccttc agcccgtcgg attaccctcc tcgtgccgtt 1500 tgctttaagc ccttccctgg caaccccctc gacaacgatc atacgccggg caagatgccg 1560 ctgacctggt ttaacatgtc ggagtttatg tgcggcaagg tcagcaactg ccttggccct 1620 gagtttaagc gcttcgacaa ctccaagacg agccgctccc cggccttcga cctggccctg 1680 gttacgcgcg tggtgtcggt cagcgatatg gagttcaagc cccacctcaa catcgacgtc 1740 aacccgtcga agggcacgat gattggcgag ttcgactgcc ccgctgacgc ctggttcttt 1800 cagggctcct gcaacgacgg ccacatgccg tacagcatcg tcatggagat cgcccttcag 1860 accagcggtg tcctcacctc cgtcctcaag gccccgctca ctatggacaa ggacgacatt 1920 ctctttcgca acctcgacgc caccgccgag atggtccgtt ccgacgtcga ttgccgcggt 1980 aagaccatca agaacttcac ccagtgcacc ggctacagca tgcttggcaa gatgggcatc 2040 caccgcttca cttttgagct ctcggtcgat gacgtcgtgt tttacaaggg ctcgaccagc 2100 tttggttggt tcacgccgga ggtgtttgag tcgcaggtcg gcctcgataa cggcaagaag 2160 gtccagccgt ggtatctgga gcagaagtcg tcgaacgtgg tgacgtacga tgtcgcctcg 2220 accgccggca aggacaagct cttctcgaag atcggctcga aggacgctca ggtccagcgt 2280 cgcaacaccc agtgcgagtt tctcgacacg atgcacatta ttccgaacac cggcaagtac 2340 aacaagggct acgcgcacgg tgagaagaag gtcaacccca acgactggtt cttctcctgc 2400 cacttttggt tcgacccggt gatgcccggc tccctcggta ttgagtccat gttccagctc 2460 atcgaggcct tttcgattga ccagggtatc gcgtccaagc atggcatcgt gaaccctacc 2520 ttcgcgcact cgaacggcaa gacctcgtgg aagtaccgcg gccagctcaa caacaagggc 2580 aagcgcatgg acagcgagat tcacatcaag gatattgtca agaacgccga cggtactgtc 2640 gatctcatcg ccgatggttt tcttctcgtg gactcgcttc gcgtttacag cgccgatgac 2700 ctccgcgtca agatcgtccc cggcactaag gctgctccca agagcgtcgc ggccgctccg 2760 cgccatgtgg ccactccgat ccccggcgtc ccctccaaca cctcctcggt ggagatctcg 2820 cttgagtccc ttaagaagga gctcctcaac ctggagaagc ccctctacct tgagacttcc 2880 aaccacatcg tcaagcagtt cggcgacgtt aacaacggcc aggcctccgt catccctccg 2940 tgcaccatta acgatctcgg tgagcgctcg tttatggaga cttacaacgt cgtcgctccc 3000 ctctacaccg gcgcgatggc gaagggcatc gcttcggcgg acctcgtcat cgctgccggt 3060 aagcgcaaga tcctcggcag cttcggcgcc ggtggcctcc cgatgcacct cgtgcgcgcc 3120 tcggtcgaga agatccaggc cgccctcccg gagggcccgt acgcggtcaa cctcatccac 3180 tcgcctttcg actcgaacct tgagaagggt aacgtggacc tctttctgga gaagggcgtc 3240 cacgtggtcg aggcctccgc ctttaccgcc ctcacgaccc aggtcgttcg ctaccgcgcc 3300 tgcggcctct cgcgtgctaa ggacggttcc gtgctgatta agaaccgcat catcggtaag 3360 gtcagccgca cggagcttgc cgagatgttc tttcgccctg ccccccagaa cctcctcgat 3420 aagctcatcg ccagcggcga gatcaccaag gagcaggcgt ccctcgctct tgaggttcct 3480 atggccgacg atgtcgctgt tgaggccgac tccggcggcc acaccgataa ccgtcccatc 3540 cacgtcatcc tcccgctgat tattaacctc cgcaaccgta tccacaagga gtgcggcttt 3600 cctgccgctc tccgcgtccg cgtcggcgct ggtggtggca tcggttgccc ctcggccgct 3660gagctccagg ttctcaagaa gggcaccatg tttccgtcgc gcgccaagaa gctctacgac 5400 ctctttaaga agtacaagtc gatcgaggag ctccctgccg acgaggtcaa gaagctggag 5460 cagaaggttt ttaagaagtc gttcgacgag gtctgggacg agactaagaa ctactacatt 5520 aaccgcctcc actcccctga gaagatcgag cgcgcggagc gtgacgccaa gctgaagatg 5580 tcgctctgct ttcgttggta cctgagcaag tcgtcccgct gggccaacac cggcgagtcg 5640 ggccgtgtcc aggactacca gatctggtgc ggccccgcca tcggctcgta caacgacttc 5700 gcgaagggct cgccctgcct tgaccctgag atccttggct cgttcccgtc ggttgtccag 5760 atcaacaagc Atattctgcg cggcgcttgc ttctaccagc gtctttcgca gctcaagtac 5820 cttaacttca actacgagga gctcgatacg ctcacctaca gcgctagcaa ctttatctaa 5880 &lt;210&gt; 122 &lt;211&gt; 4368 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Codon-optimized PFA3 &lt;;400&gt; 122 atggttggcc tgcagatgaa gaagaagcct gtgtgggaga tgtcgaagga ggagcagtcg 60 tccggcaaga acgtcgtctt tgactacgac gagctcctcg aattcgcgga gggtgacatc 120 ggcaaggtgt tcggccccaa gtttgacatc atcgacaagt acagccgccg tgtgc gcctc 180 ccggcccgcg agtacctcct cgtcacccgt gtcacgctca tggatgccga ggtcggcaac 240 ttccgcgtcg gctcgcgcat ggtcaccgag tacgacgtcc cggtgaacgg cgagctttcc 300 cagggcggcg acgttccctg ggccgtcctc gtcgagtcgg gccagtgcga cctcatgctt 360 atctcgtaca tgggcattga ctttcagtgc aagggtgacc gcgtttaccg ccttctcaac 420 acgaccctca cgttctacgg tgtcgcccac gagggcgaga ctctcgttta cgacatccgc 480 gtcactggtt tcgccaaggg catgcacggc gagattagca tgttcttctt cgagtacgac 540 tgctacgtca acggccgcct gctcatcgag atgcgcgacg gttgcgctgg cttcttcacg 600 gacgaggagc tcgccgcggg caagggcgtc atcaagaccg tcgctgagct ccacaagcgc 660 aagtcgattg tgcccaagtc gatcaagcct tttgccctca accccgccgt ccacaagacg 720 atgttcagcg agaacgacat ggagaagctt tgcgagcgcc agtgggagaa cgtcctcggc 780 tccggcctcc agggcatcga ctacaagctg tgcgcccgca agatgctcat gatcgaccgc 840 atcacgaaga tccagcacaa cggcggtgcg tacggcctcg gcctcctcgt tggcgagaag 900 attcttgagc gcgaccattg gtacttccct tgccacttcg tcaaggacca ggtgatggcg 960 ggctccctcg ttagcgacgg ctgctcgcag ctgctcaagc tttacatgct ttggctcggc 1020 ctccacgacg tggtccccga tttccagttc cgtcctgtcc ctggccagcc caacaaggtg 1080 cgctgccgtg gccagatcag cccccatcgt ggcaagctcg tgtacgtgat ggagattcgc 1140 gagatgggtt tcaacgagtc caccggccag ccctacgcga tcgctgacgt tgacattatc 1200 gatgtgaact acgagctcgg ccagtccttt gacatggccg acatcgactc gtacggccgt 1260 ggcaacctct ccaagaagat tgtcgtcgat ttcaagggca ttgcgctcca gatggagggc 1320 accgtcaaga gctccaacat catcgattcg tcccccaagt ccacgattat ccagccgccg 1380 cccaactgcc tccgcggcga tcctctcgcc ccctcgcagg tcacctggca cccgatggcc 1440 203 201038734 ggtgtcaacg gcgcccccgc cccctccttc agcccgtcgg attaccctcc tcgtgccgtt 1500 tgctttaagc ccttccctgg caaccccctc gacaacgatc atacgccggg caagatgccg 1560 ctgacctggt ttaacatgtc ggagtttatg tgcggcaagg tcagcaactg ccttggccct 1620 gagtttaagc gcttcgacaa ctccaagacg agccgctccc cggccttcga cctggccctg 1680 gttacgcgcg tggtgtcggt cagcgatatg gagttcaagc cccacctcaa catcgacgtc 1740 aacccgtcga agggcacgat gattggcgag ttcgactgcc ccgctgacgc ctggttcttt 1800 cagggctcct gcaacgacgg ccacatgccg tacagcatcg t catggagat cgcccttcag 1860 accagcggtg tcctcacctc cgtcctcaag gccccgctca ctatggacaa ggacgacatt 1920 ctctttcgca acctcgacgc caccgccgag atggtccgtt ccgacgtcga 1980 aagaccatca agaacttcac ccagtgcacc ggctacagca tgcttggcaa gatgggcatc 2040 accgccggca aggacaagct cttctcgaag ttgccgcggt caccgcttca cttttgagct ctcggtcgat gacgtcgtgt tttacaaggg ctcgaccagc 2100 tttggttggt tcacgccgga ggtgtttgag tcgcaggtcg gcctcgataa cggcaagaag 2160 gtccagccgt ggtatctgga gcagaagtcg tcgaacgtgg tgacgtacga tgtcgcctcg 2220 atcggctcga aggacgctca ggtccagcgt 2280 cgcaacaccc agtgcgagtt tctcgacacg atgcacatta ttccgaacac cggcaagtac 2340 aacaagggct acgcgcacgg tgagaagaag gtcaacccca acgactggtt cttctcctgc 2400 cacttttggt tcgacccggt gatgcccggc tccctcggta ttgagtccat gttccagctc 2460 atcgaggcct tttcgattga ccagggtatc gcgtccaagc atggcatcgt gaaccctacc 2520 ttcgcgcact cgaacggcaa gacctcgtgg aagtaccgcg gccagctcaa caacaagggc 2580 aagcgcatgg acagcgagat tcacatcaag gatattgtca agaacgccga cggtactgtc 2640 gatctcatcg ccgatggttt tcttctcgtg gact cgcttc gcgtttacag cgccgatgac 2700 ctccgcgtca agatcgtccc cggcactaag gctgctccca agagcgtcgc ggccgctccg 2760 cgccatgtgg ccactccgat ccccggcgtc ccctccaaca cctcctcggt ggagatctcg 2820 cttgagtccc ttaagaagga gctcctcaac ctggagaagc ccctctacct tgagacttcc 2880 aaccacatcg tcaagcagtt cggcgacgtt aacaacggcc aggcctccgt catccctccg 2940 tgcaccatta acgatctcgg tgagcgctcg tttatggaga cttacaacgt cgtcgctccc 3000 ctctacaccg gcgcgatggc gaagggcatc gcttcggcgg acctcgtcat cgctgccggt 3060 aagcgcaaga tcctcggcag cttcggcgcc ggtggcctcc cgatgcacct cgtgcgcgcc 3120 tcggtcgaga agatccaggc cgccctcccg gagggcccgt acgcggtcaa cctcatccac 3180 tcgcctttcg actcgaacct tgagaagggt aacgtggacc tctttctgga gaagggcgtc 3240 cacgtggtcg aggcctccgc ctttaccgcc ctcacgaccc aggtcgttcg ctaccgcgcc 3300 tgcggcctct cgcgtgctaa ggacggttcc gtgctgatta agaaccgcat catcggtaag 3360 gtcagccgca cggagcttgc cgagatgttc tttcgccctg ccccccagaa cctcctcgat 3420 aagctcatcg ccagcggcga gatcaccaag gagcaggcgt ccctcgctct tgaggttcct 3480 atggccgacg atgtcgctgt tgaggcc Gac tccggcggcc acaccgataa ccgtcccatc 3540 cacgtcatcc tcccgctgat tattaacctc cgcaaccgta tccacaagga gtgcggcttt 3600 cctgccgctc tccgcgtccg cgtcggcgct ggtggtggca tcggttgccc ctcggccgct 3660

204 201038734 gtcgccgcct tcaacatggg cgcggccttc ctgatcaccg gctccgttaa ccaggtgagc 3720 aagcagtccg gcacgtgcga cattgtgcgc aagcagctta gcgaggccag ctactccgac 3780 atcacgatgg ctcccgccgc tgacatgttc gaccagggcg tggagctcca ggtcctcaag 3840 aagggtacga tgtttccctc gcgcgccaag aagctctacg agctcttttg catgtacaac 3900 agctttgacg acatgccgaa gtccgagctc cagcgcctgg agaagcgcat tttccagaag 3960 agcctcgccg aggtctggga ggagactaag gacttttaca tcaaccgcct caacaacccg 4020 gagaagatcg agcacgccga gaagaaggac cccaagctca agatgtccct ttgctttcgc 4080 tggtatctcg gcctttcgag cttttgggcc aacaacggca tcaaggagcg cagcatggat 4140 taccagattt ggtgcggccc ggccattggc agctacaacg acttcgtgaa gggcacctac 4200 ctcgaccccg ccgtcgccgg ttcgtacccc tgcgtggtcc agatcaacat gcagatcctc 4260 cgcggtgcgt gcttcctcca gcgcgtccgc gccattaagc acgacccgcg cctcgatatc 4320 gacgttgatg aggacgtctt tacctaccgc cccgagagca ccctctaa 4368 &lt;210&gt; &lt;211〉 &lt;212〉 &lt;213&gt; Ο 3 A工 2 3n&gt; 1 3 D Λ 列 &lt;220&gt; &lt;223&gt; 合成引子pDS233 &gt; &gt; &gt; &gt; 0 12 3 2 2 2 2 2 2 2 2 &lt; &lt; &lt; &lt; 其他特徵 (25)·(25) η 為 a ' c、g，或 t &gt; &gt; &gt; &gt; 0 12 3 2 2 2 2 2 2 2 2 &lt; &lt; &lt; &lt; 其他特徵 (28).. (28) n為a、c、g，或 t Ο &lt;221〉其他特徵 &lt;222〉 （31)..(31) &lt;223〉 y 為 c 或 t &lt;400&gt; 123 tgatatggga ggaatgaatt gtgtngtnga ygc 33 &lt;210&gt; &lt;211&gt; &lt;212&gt; &lt;213&gt; &lt;220&gt; &lt;223〉 124 36 DNA 人工序列 合成引子pDS235 &lt;220&gt; &lt;221&gt; &lt;222&gt; &lt;223&gt; &lt;220&gt; &lt;221&gt; &lt;222&gt; &lt;223&gt; &lt;220&gt; &lt;221〉 &lt;222&gt; 其他特徵 (28) · , (28) η 為 a、c、g，或 t 其他特徵 (31)..(31) r為a或g 其他特徵 (34)..(34) 205 201038734 &lt;223&gt; η 為 a、c、g，或 t &lt;400&gt; 124 ttccataaca aaatgataat tagctccncc raancc &lt;210&gt; 125 &lt;211〉 23 &lt;212&gt; DNA &lt;213&gt; 人工序列 &lt;220&gt; &lt;223&gt; 合成引子prDS183 &lt;22〇&gt; &lt;221&gt; 其他特徵 &lt;222&gt; (15)..(15) &lt;223〉 y為c或t &lt;220&gt; &lt;221〉 其他特徵 &lt;222&gt; (19) .. (19) &lt;223&gt; m為a或c &gt; &gt; &gt; &gt; 0 12 3 2 2 2 2 2 2 2 2 &lt; &lt; &lt; &lt; 其他特徵 (21) . · (21) η 為a、c、g，或t &lt;400&gt; 125 ggcggccaca ccgayaaymg ncc &lt;210&gt; 126 &lt;211&gt; 22 &lt;212&gt; DNA &lt;213〉 人工序列 &lt;22〇&gt; &lt;223&gt; 合成引子prDS184 &lt;220&gt; &lt;221&gt; 其他特徵 &lt;222&gt; (14)..(14) &lt;223&gt; η 為a、c、g，或t &lt;220&gt; &lt;221〉 其他特徵 &lt;222&gt; (20) .. (20) &lt;223&gt; r 為 a 或 g &lt;40〇&gt; 126 cggggccgca ccanayytgr ta &lt;210&gt; 127 &lt;211&gt; 17 &lt;212&gt; DNA &lt;213&gt; 人工序列 &lt;220&gt; &lt;223&gt; 合成引子prDS181 &gt; &gt; &gt; &gt; 0 12 3 2 2 2 2 2 2 2 2 &lt; &lt; &lt; &lt; 其他特徵 (9)..(9) η 為 a、c、g ,或 t &lt;220&gt; &lt;221&gt; 其他特徵 &lt;222&gt; (12)..(12) &lt;223&gt; η為a、c、g，或 t 206 &lt;220&gt; 201038734 &gt; &gt; &gt; 12 3 2 2 2 2 2 2 &lt; &lt; &lt; 其他特徵 (14)..(14) s為c或g &lt;220&gt; &lt;221&gt; 其他特徵 &lt;222&gt; (15)..(15) &lt;223&gt; η 為 a、c、g，或 t &lt;400&gt; 127 17 tccttcggng cngsngg Ο ο 207204 201038734 gtcgccgcct tcaacatggg cgcggccttc ctgatcaccg gctccgttaa ccaggtgagc 3720 aagcagtccg gcacgtgcga cattgtgcgc aagcagctta gcgaggccag ctactccgac 3780 atcacgatgg ctcccgccgc tgacatgttc gaccagggcg tggagctcca ggtcctcaag 3840 aagggtacga tgtttccctc gcgcgccaag aagctctacg agctcttttg catgtacaac agatgtccct ttgctttcgc 3900 agctttgacg acatgccgaa gtccgagctc cagcgcctgg agaagcgcat tttccagaag 3960 agcctcgccg aggtctggga ggagactaag gacttttaca tcaaccgcct caacaacccg 4020 gagaagatcg agcacgccga gaagaaggac cccaagctca 4080 tggtatctcg gcctttcgag cttttgggcc aacaacggca tcaaggagcg cagcatggat 4140 taccagattt ggtgcggccc ggccattggc agctacaacg acttcgtgaa gggcacctac 4200 ctcgaccccg ccgtcgccgg ttcgtacccc tgcgtggtcc agatcaacat gcagatcctc 4260 cgcggtgcgt gcttcctcca gcgcgtccgc gccattaagc acgacccgcg cctcgatatc 4320 gacgttgatg aggacgtctt tacctaccgc cccgagagca ccctctaa 4368 &lt; 210 &gt; &lt; 211> &lt; 212> &lt; 213 &gt; Ο 3 A worker 2 3n&gt; 1 3 D Λ column &lt;220&gt;&lt;223&gt; Synthetic primer pDS233 &gt;&gt;&gt;&gt; 0 12 3 2 2 2 2 2 2 2 2 &lt;&lt;&lt;&lt; Other characteristics (25)·(25) η is a ' c, g, or t &gt;&gt;&gt;&gt; 0 12 3 2 2 2 2 2 2 2 2 &lt;&lt;&lt;&lt; Other characteristics (28).. (28) n is a, c, g, or t Ο &lt;221> Other features &lt;222> (31 )..(31) &lt;223> y is c or t &lt;400&gt; 123 tgatatggga ggaatgaatt gtgtngtnga ygc 33 &lt;210&gt;&lt;211&gt;&lt;212&gt;&lt;213&gt;&lt;220&gt;&lt;223> 124 36 DNA artificial sequence synthesis primer pDS235 &lt;220&gt;&lt;221&gt;&lt;222&gt;&lt;223&gt;&lt;220&gt;&lt;221&gt;&lt;222&gt;&lt;223&gt;&lt;220&gt;&lt;221〉&lt;222&gt; Other features (28) · , (28) η is a, c, g, or t Other features (31).. (31) r is a or g Other features (34)..(34) 205 201038734 &lt;223&gt; η is a, c, g, or t &lt;400&gt; 124 ttccataaca aaatgataat tagctccncc raancc &lt;210&gt; 125 &lt;211> 23 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Synthetic primer prDS183 &lt;22〇&gt;&lt;221&gt; Other features &lt;222&gt; (15)..(15) &lt;223> y Is c or t &lt;220&gt;&lt;221> Other features &lt;222&gt; (19) .. (19) &lt;223&gt; m is a or c &gt;&gt;&gt;&gt; 0 12 3 2 2 2 2 2 2 2 2 &lt;&lt;&lt;&lt; Other characteristics (21) . (21) η is a, c, g, or t &lt;400&gt; 125 ggcggccaca ccgayaaymg ncc &lt;210&gt; 126 &lt;211&gt;&lt;212&gt; DNA &lt;213> Artificial sequence &lt;22〇&gt;&lt;223&gt; Synthesis primer prDS184 &lt;220&gt;&lt;221&gt; Other characteristics &lt;222&gt; (14)..(14) &lt;223&gt; η is a, c, g, or t &lt;220&gt;&lt;221> Other features &lt;222&gt; (20) .. (20) &lt;223&gt; r is a or g &lt;40〇&gt; 126 cggggccgca ccanayytgr Ta &lt;210&gt; 127 &lt;211&gt; 17 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Synthesis primer prDS181 &gt;&gt;&gt;&gt; 0 12 3 2 2 2 2 2 2 2 2 &lt;&lt;&lt;&lt; Other characteristics (9)..(9) η is a, c, g, or t &lt;220&gt;&lt;221&gt; Other characteristics &lt;222&gt; (12)..( 12) &lt;223&gt; η is a, c, g, or t 206 &lt;220&gt; 201038734 &gt;&gt;&gt; 12 3 2 2 2 2 2 2 &lt;&lt;&lt;&lt; (14)..(14) s is c or g &lt;220&gt;&lt;221&gt; Other characteristics &lt;222&gt; (15)..(15) &lt;223&gt; η is a, c, g, or t &lt;;400&gt; 127 17 tccttcggng cngsngg Ο ο 207

Claims (1)

201038734 七、申請專利範圍： 1. 一種經單離的核酸分子，其係選自於以下所構成的群 組： (a) —核酸分子，其包含與序列辨識編號：1有至少 80%的同一性之多核苷酸序列，其中該多核苷酸序列編 碼一多肽，該多肽包含選自於以下所構成的群組之多不 飽和脂肪酸(PUFA)合成酶活性：β-酮基醯基-ACP合成酶 (KS)活性、丙二醯基-CoA : ACP醯基轉移酶(MAT)活 性、醯基載體蛋白質（ACP)活性、酮還原酶(KR)活性、β-羥基醯基-ACP脫水酶(DH)活性，以及其等之組合； (b) —核酸分子，其包含與序列辨識編號：7有至少 80%的同一性之多核苷酸序列，其中該多核苷酸序列編 碼一多肽，其包含KS活性； (c) 一核酸分子，其包含與序列辨識編號：9有至少 80%的同一性之多核苷酸序列，其中該多核苷酸序列編 碼一多肽，其包含MAT活性； (d) —核酸分子，其包含與序列辨識編號：13、15、 17、19、21，或23的任一者有至少80%的同一性之多核 苷酸序列，其中該多核苷酸序列編碼一多肽，其包含 ACP活性； (e) —核酸分子，其包含與序列辨識編號：11有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含ACP活性； (f) 一核酸分子，其包含與序列辨識編號：25有至 1 201038734 少80%的同—性之多核《序列，其中該多核苦酸序列 編碼一多肽，其包含KR活性；以及 (g) —核酸分子，其包含與序列辨識編號· 27有至 少80%的同—性之多核該序列，其中該多核魏序列 編碼一多肽，其包含DH活性。 2·如申請專利範圍第1項之經單離的核酸分子，其中該等 多核苦酸序列係與序列辨識編號：1、7、9、u、13、 15、17、19、21、23、25 ’ 以及27有至少90%同一性。 3.如申凊專利範圍第1項之經單離的核酸分子，其中該等 多核苷酸序列係與序列辨識編號：1、7、9、丨丨、13、 15、17、19、21、23、25,以及27各別有至少95%同一 性。 4_如申請專利範圍第丨項之經單離的核酸分子其中該等 核酸分子各別包含序列辨識編號：1、7、9、u、13、 15、17、19、2卜23、25 ’及27之多核苷酸序列。 5. —種經單離的核酸分子，其係選自於以下所構成的群 組： (a) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：2有至少8〇%的 同一性之胺基酸序列，以及其中該多肽包含選自於以下 所構成的群組之PUFA合成酶活性：KS活性、MAT活性、 ACP活性、KR活性、DH活性 '以及其等之組合； (b) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：8有至少8〇%的 201038734 同丨生之胺基酸序列，以及其中該多肽包含ks活性； (e)—核酸分子，其包含編碼一多肽之多核苷酸序 列’其中該多肽包含與序列辨識編號：1〇有至少8〇%的 同丨生之胺基酸序列’以及其中該多肽包含MAT活性； 酸分子，其包含編碼一多肽之多核苷酸序 肽包含與序列辨識編號：I4、16、I8、2〇， 一性之胺基酸序列，以201038734 VII. Patent Application Range: 1. An isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising at least 80% identical to the sequence identification number: A polynucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising a polyunsaturated fatty acid (PUFA) synthase activity selected from the group consisting of: β-ketothiol-ACP Synthetase (KS) activity, propylenediyl-CoA: ACP thiol transferase (MAT) activity, thiol-based carrier protein (ACP) activity, ketoreductase (KR) activity, β-hydroxy thiol-ACP dehydratase (DH) activity, and combinations thereof; (b) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 7, wherein the polynucleotide sequence encodes a polypeptide, It comprises KS activity; (c) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 9, wherein the polynucleotide sequence encodes a polypeptide comprising MAT activity; d) - Nucleic acid molecule, which contains and sequence identification number: 13, 1 5. The polynucleotide of any of 5, 17, 19, 21, or 23 having at least 80% identity, wherein the polynucleotide sequence encodes a polypeptide comprising ACP activity; (e) a nucleic acid molecule, It comprises a polynucleotide sequence having at least 80% identity to sequence identification number: 11, wherein the polynucleotide sequence encodes a polypeptide comprising ACP activity; (f) a nucleic acid molecule comprising the sequence identification number :25至至201038734 80% less homo-nuclear "sequence, wherein the polynucleotide sequence encodes a polypeptide comprising KR activity; and (g) - a nucleic acid molecule comprising the sequence identification number. There is at least 80% homozygous multinuclear sequence, wherein the multinuclear Wei sequence encodes a polypeptide comprising DH activity. 2. The isolated nucleic acid molecule of claim 1, wherein the polynucleic acid sequence and sequence identification number: 1, 7, 9, u, 13, 15, 17, 19, 21, 23, 25 ' and 27 have at least 90% identity. 3. The isolated nucleic acid molecule of claim 1, wherein the polynucleotide sequence and sequence identification number: 1, 7, 9, 丨丨, 13, 15, 17, 19, 21, 23, 25, and 27 each have at least 95% identity. 4) The isolated nucleic acid molecule of claim 3, wherein the nucleic acid molecules each comprise a sequence identification number: 1, 7, 9, u, 13, 15, 17, 19, 2, 23, 25' And a polynucleotide sequence of 27. 5. An isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises a sequence identification number : 2 an amino acid sequence having at least 8 % identity, and wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of KS activity, MAT activity, ACP activity, KR activity, DH activity And a combination thereof; (b) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 8% of the amino acid of 201038734 with a sequence identification number: 8 a sequence, and wherein the polypeptide comprises ks activity; (e) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide wherein the polypeptide comprises at least 8% of the same number as the sequence identification number: An amino acid sequence 'and wherein the polypeptide comprises MAT activity; an acid molecule comprising a polynucleotide sequence encoding a polypeptide comprising the sequence identification number: I4, 16, I8, 2〇, a monobasic amino acid sequence To 及其中该多肽包含ACP活性； (e) ~核酸分子，其包含編碼一多肽之多核苷酸序 列’其中該多肽包含與序列辨識編號：12有至少8〇0/〇的 同一性之胺基酸序列，以及其中該多肽包含ACP活性； ⑺一核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：26有至少80%的 同一性之胺基酸序列，及其中該多肽包含KR活性；以 及And wherein the polypeptide comprises ACP activity; (e) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino group having at least 8 〇 0/〇 identity with sequence number: 12 An acid sequence, and wherein the polypeptide comprises ACP activity; (7) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 26. And wherein the polypeptide comprises KR activity; (d)—核 列，其中該多 或24的任何—者有至少80%的同 (g)—核酸分子，其包含編碼一多肽之多核苷酸序 列’其中該多肽包含與序列辨識編號：28有至少80%的 同一性之胺基酸序列，以及其中該多肽包含DH活性。 6_如申清專利範圍第5項之經單離的核酸分子，其中該等 胺基酸序列與序列辨識編號：2、8、1〇、12、14、16、 18、20、22、24、26，及28各別有至少90%的同一性。 7.如申請專利範圍第5項之經單離的核酸分子，其中該等 胺基酸序列與序列辨識編號：2、8、1〇、12、14、16、 18、20、22、24、26，及28各別有至少95%的同一性。 3 201038734 8. 如申請專利範圍第5項之經單離的核酸分子，其中該等 多肽各別包含序列辨識編號：2、8、10、12、14、16、 18、20、22、24、26，及28之胺基酸序歹ij ° 9. 一種經單離的核酸分子，其係選自於以下所構成的群 組： (a) —核酸分子，其包含與序列辨識編號：3有至少 80%的同一性之多核苷酸序列，其中該多核苷酸序列編 碼一多肽，其包含選自於以下所構成的群組之PUFA合 成酶活性：KS活性、鏈長度因子（CLF)活性、醯基轉移 酶(AT)活性、烯醯基-ACP還原酶(ER)活性，以及其等之 組合； (b) —核酸分子，其包含與序列辨識編號：29有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含KS活性； (c) 一核酸分子，其包含與序列辨識編號：31有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含CLF活性； (d) —核酸分子，其包含與序列辨識編號：33有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含AT活性；以及 (e) —核酸分子，其包含與序列辨識編號：35有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含ER活性。 10. 如申請專利範圍第9項之經單離的核酸分子，其中該等 4 201038734 多核苷酸序列係各別與序列辨識編號：3、29、31、33， 及35有至少90%的同一性。 11. 如申請專利範圍第9項之經單離的核酸分子，其中該等 多核苷酸序列係各別與序列辨識編號：3、29、31、33， 及35有至少95%的同一性。 12. 如申請專利範圍第9項之經單離的核酸分子，其中該等 核酸分子各別包含序列辨識編號：3、29、31、33，以 及35的多核苷酸序列。 13. —種經單離的核酸分子，其係選自於以下所構成的群 組： (a) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：4有至少80%的 同一性之胺基酸序列，以及其中該多肽包含選自於以下 所構成的群組之PUFA合成酶活性：KS活性、CLF活性、 AT活性、ER活性，及其等之組合； (b) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：30有至少80%的 同一性之胺基酸序列，以及其中該多肽包含KS活性； (c) 一核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：32有至少80%的 同一性之胺基酸序列，以及其中該多肽包含CLF活性； (d) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：34有至少80%的同 一性之胺基酸序列，以及其中該多肽包含AT活性；以及 5 201038734 (e) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：36有至少80%的 同一性之胺基酸序列，以及其中該多肽包含ER活性。 14. 如申請專利範圍第13項之經單離的核酸分子，其中該等 胺基酸序列係各別與序列辨識編號：4、30、32、34， 及36有至少90%的同一性。 15. 如申請專利範圍第13項之經單離的核酸分子，其中該等 胺基酸序列係各別與序列辨識編號：4、30、32、34， 及36有至少95%的同一性。 16. 如申請專利範圍第13項之經單離的核酸分子，其中該等 多肽各別包含序列辨識編號：4、30、32、34，及36之 胺基酸序列。 17. —種經單離的核酸分子，其係選自於以下所構成的群 組： (a) —核酸分子，其包含與序列辨識編號：5有至少 80%的同一性之多核苷酸序列，其中該多核苷酸序列編 碼一多肽，其包含選自於以下所構成的群組之PUFA合 成酶活性：DH活性、ER活性、以及其等之組合； (b) —核酸分子，其包含與序列辨識編號：37有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含DH活性； (c) 一核酸分子，其包含與序列辨識編號：39有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含DH活性；以及 201038734 (d) —核酸分子，其包含與序列辨識編號：41有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含ER活性。 18. 如申請專利範圍第17項之經單離的核酸分子，其中該等 多核苷酸序列係各別與序列辨識編號：5、37、39，及 41有至少90%的同一性。 19. 如申請專利範圍第17項之經單離的核酸分子，其中該等 多核苷酸序列係各別與序列辨識編號：5、37、39，及 41有至少95%的同一性。 20. 如申請專利範圍第17項之經單離的核酸分子，其中該等 核酸分子各別包含序列辨識編號：5、37、39，及41的 多核苷酸序列。 21. —種經單離的核酸分子，其係選自於以下所構成的群 組： (a) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：6有至少80%的 同一性之胺基酸序列，以及其中該多肽包含選自於以下 所構成的群組之PUFA合成酶活性·· DH活性、ER活性、 及其等之組合； (b) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：38有至少80%的 同一性之胺基酸序列，以及其中該多肽包含DH活性； (c) 一核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：40有至少80%的 7 201038734 同一性之胺基酸序列，以及其中該多肽包含DH活性； 以及 (d) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：42有至少80%的 同一性之胺基酸序列，以及其中該多肽包含ER活性。 22. 如申請專利範圍第21項之經單離的核酸分子，其中該等 胺基酸序列係各別與序列辨識編號：6、38、40，及42 有至少90%的同一性。 23. 如申請專利範圍第21項之經單離的核酸分子，其中該等 胺基酸序列係各別與序列辨識編號：6、38、40，及42 有至少95%的同一性。 24. 如申請專利範圍第21項之經單離的核酸分子，其中該等 多肽各別包含序列辨識編號：6、38、40，及42之胺基 酸序列。 25. —種經單離的核酸分子，其包含編碼一多肽之多核苷酸 序列，該多肽包含選自於以下所構成的群組之PUFA合 成酶活性：KS活性、MAT活性、ACP活性、KR活性、 CLF活性、AT活性、ER活性、DH活性、以及其等之組 合， 其中該多核苷酸係於嚴苛條件下與申請專利範圍 第1-24項中任一項之多核苷酸序列之互補物雜交。 26. —種經單離的核酸分子，其包含一多核苷酸序列，其係 與申請專利範圍第1-24項中任一項之多核苷酸序列為 完全互補的。 201038734 27. —種重組型核酸分子，其包含如申請專利範圍第1-25項 中任一項之核酸分子或其等之組合以及一轉錄控制序 列。 28. 如申請專利範圍第27項之重組型核酸分子，其中該重組 型核酸分子為一重組型載體。 29. —種宿主細胞，其表現如申請專利範圍第1-25項中任一 項之核酸分子、如申請專利範圍第27或28項之重組型核 酸分子，或其等之組合。 30. 如申請專利範圍第29項之宿主細胞，其中該宿主細胞係 選自於以下所構成的群組：植物細胞、微生物細胞，以 及動物細胞。 31. 如申請專利範圍第30項之宿主細胞，其中該宿主細胞係 微生物細胞。 32. 如申請專利範圍第31項之微生物細胞，其中該微生物細 胞係一細菌。 33. 如申請專利範圍第31項之微生物細胞，其中該微生物細 胞為破囊壺菌。 34. 如申請專利範圍第33項之微生物細胞，其中該破囊壺菌 為裂殖壺菌屬（Schizochytrium)或破嚢壺菌屬 (Thraustochytrium) ° 35. —種產生至少一個PUFA的方法，其包含： 於有效產生PUFA的條件下於一宿主細胞中表現 PUFA合成酶基因， 其中該PUFA合成酶基因包含如申請專利範圍第 9 201038734 36. 37. 38. 39. 40. 41. 42. 1-25項中任一項之經單離的核酸分子、如申請專利範圍 第27或28項之重組型核酸分子，或其等之組合，以及 其中至少一個PUFA係被生產。 如申請專利範圍第3 5項之方法，其中該宿主細胞係選自 於以下所構成的群組：植物細胞、經單離的動物細胞， 以及微生物細胞。 如申請專利範圍第35項之方法，其中該至少一個PIJFA 包含二十二碳六烯酸(DHA)。 一種生產富含DHA的脂質的方法，其包含： 於有效產生脂質的條件下於一宿主細胞中表現 PUFA合成酶基因，其中該PUFA合成酶基因包含如申請 專利範圍第1-25項中任一項之經單離的核酸分子、如申 請專利範圍第27或28項之重組型核酸分子，或其等之組 合於該宿主細胞中，以及 其中富含DHA的脂質被生產。 一種製造重組型載體的方法’其包含將如申請專利範圍 第1-25項中任一項之經單離的核酸分子插入至—载體 内。 一種製造重組塑宿主細胞的方法，其包含將如申請專利 範圍第39項之重組型載體引入至一宿主細胞中。 如申請專利範圍第4 〇項之方法，其中該宿主細胞係選自 於以下所構成的群組：植物細胞、經單離的動物細胞， 以及微生物細胞。 —種經單離的多肽，其係由申請專利範圍第項中任 201038734 一項之多核苷酸序列所編碼。 43. —種經單離的多肽，其係選自於以下所構成的群組： (a) —多肽，其包含與序列辨識編號：2有至少80% 的同一性之胺基酸序列，其中該多肽包含選自於以下所 構成的群組之PUFA合成酶活性：KS活性、MAT活性、 ACP活性、KR活性、DH活性，及其等之組合； (b) —多肽，其包含與序列辨識編號：8有至少80% 的同一性之胺基酸序列，其中該多肽包含KS活性； (c) 一多肽，其包含與序列辨識編號：10有至少80% 的同一性之胺基酸序列，其中該多肽包含MAT活性； (d) —多肽，其包含與序列辨識編號：14、16、18、 20、22，或24的任何一者有至少80%的同一性之胺基酸 序列，其中該多肽包含ACP活性； (e) —多肽，其包含與序列辨識編號：12有至少80% 的同一性之胺基酸序列，其中該多肽包含ACP活性； (f) 一多肽，其包含與序列辨識編號：26有至少80% 的同一性之胺基酸序列，其中該多肽包含KR活性；以 及 (g) —多肽，其包含與序列辨識編號：28有至少80% 的同一性之胺基酸序列，其中該多肽包含DH活性。 44. 如申請專利範圍第43項之經單離的多肽，其中該等胺基 酸序列各別與序列辨識編號：2、8、10、12、14、16、 18、20、22、24、26，及28有至少 90%的同一性。 45. 如申請專利範圍第43項之經單離的多肽，其中該等胺基 11 201038734 酸序列各別與序列辨識編號：2、8、10、12、14、16、 18、20、22、24、26，及28有至少 95%的同一性。 46. 如申請專利範圍第43項之經單離的多肽，其中該等多肽 各別包含序列辨識編號：2、8、10、12、14、16、18、 20、22、24、26，及28之胺基酸序列。 47. —種經單離的多肽，其係選自於以下所構成的群組： (a) —多肽，其包含與序列辨識編號：4有至少80% 的同一性之胺基酸序列，其中該多肽包含選自於以下所 構成的群組之PUFA合成酶活性：KS活性、CLF活性、 AT活性、ER活性、以及其等之組合； (b) —多肽，其包含與序列辨識編號：30有至少80% 的同一性之胺基酸序列，其中該多肽包含KS活性； (c) 一多肽，其包含與序列辨識編號：32有至少80% 的同一性之胺基酸序列，其中該多肽包含CLF活性； (d) —多肽，其包含與序列辨識編號：34有至少80% 的同一性之胺基酸序列，其中該多肽包含AT活性；以及 (e) —多肽，其包含與序列辨識編號：36有至少80% 的同一性之胺基酸序列，其中該多肽包含ER活性。 48. 如申請專利範圍第47項之經單離的多肽，其中該等胺基 酸序列係各別與序列辨識編號：4、30、32、34，及36 有至少90°/。的同一性。 49. 如申請專利範圍第47項之經單離的多肽，其中該等胺基 酸序列係各別與序列辨識編號：4、30、32、34，及36 有至少95%的同一性。 12 201038734 50. 如申請專利範圍第47項之經單離的多肽，其中該等多肽 各別包含序列辨識編號：4、30、32、34，及36的胺基 酸序列。 51. —種經單離的多肽，其係選自於以下所構成的群組： (a) —多肽，其包含與序列辨識編號：6有至少80% 的同一性之胺基酸序列，其中該多肽包含選自於以下所 構成的群組之PUFA合成酶活性：DH活性、ER活性、以 及其等之組合； ^ (b) —多肽，其包含與序列辨識編號：38有至少80% 的同一性之胺基酸序列，其中該多肽包含DH活性； (c) 一多肽，其包含與序列辨識編號：40有至少80% 、 的同一性之胺基酸序列，其中該多肽包含DH活性；以 -— 及 (d) —多肽，其包含與序列辨識編號：42有至少80% 的同一性之胺基酸序列，其中該多肽包含ER活性。 52. 如申請專利範圍第51項之經單離的多肽，其中該等胺基 ❹ 酸序列係各別與序列辨識編號：6、38、40，及42有至 少90%的同一性。 53. 如申請專利範圍第51項之經單離的多肽，其中該等胺基 酸序列係各別與序列辨識編號：6、38、40，及42有至 少95%的同一性。 54. 如申請專利範圍第51項之經單離的多肽，其中該等多肽 各別包含序列辨識編號：6、38、40，及42之胺基酸序 列。 13 201038734 55. 如申請專利範圍第42-54項中任一項之經單離的多肽， 其中該多肽為一融合多肽。 56. —種組成物，其包含如申請專利範圍第42-54項中任一 項之多肽以及生物上可接受的載體。 57. —種於具有PUFA合成酶活性的有機體内增加DHA的生 產之方法，其包含： 於有效產生DHA的條件下於該有機體中表現如申 請專利範圍第1-25項中任一項之經單離的核酸分子、如 申請專利範圍第27或28項之重組型核酸分子，或其等之 組合， 其中該PUFA合成酶活性於該有機體中取代不活化 或被刪除的活性、引入新的活性、或提高現存的活性， 以及 其中該有機體中的DHA之生產為增加的。 58. —種自宿主細胞單離脂質的方法，其包含： (a) 於有效產生脂質的條件下於該宿主細胞中表現 PUFA合成酶基因，其中PKS系統包含如申請專利範圍第 1-25項中任一項之經單離的核酸分子、如申請專利範圍 第27或28項之重組型核酸分子，或其等之組合於該宿主 細胞中，以及 (b) 自該宿主細胞單離脂質。 59. 如申請專利範圍第58項之方法，其中該宿主細胞係選自 於以下所構成的群組：植物細胞、經單離的動物細胞， 以及微生物細胞。 14 201038734 60. 如申請專利範圍第58項之方法，其中該等脂質包含 DHA。 61. —種經單離的核酸分子，其係選自於以下所構成的群 組： (a) —核酸分子，其包含與序列辨識編號：68或序 列辨識編號：120有至少80%的同一性之多核苷酸序 列，其中該多核苷酸序列編碼一多肽，其包含選自於以 下所構成的群組之PUFA合成酶活性：KS活性、MAT活 性、ACP活性、KR活性、DH活性、以及其等之組合； (b) —核酸分子，其包含與序列辨識編號：74有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含KS活性； (c) 一核酸分子，其包含與序列辨識編號：76有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含MAT活性； (d) —核酸分子，其包含與序列辨識編號：80、82、 84、86、88、90、92、94、96或98的任一者有至少 80% 的同一性之多核苷酸序列，其中該多核苷酸序列編碼一 多肽，其包含ACP活性； (e) —核酸分子，其包含與序列辨識編號：78有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含ACP活性； (f) 一核酸分子，其包含與序列辨識編號：100有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 15 201038734 編碼一多肽，其包含KR活性；以及 (g) —核酸分子，其包含與序列辨識編號：118有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含DH活性。 62. 如申請專利範圍第61項之經單離的核酸分子，其中該等 多核苷酸序列係各別與序列辨識編號：68、74、76、78、 80、82、84、86、88、90、92、94、96、98、100、118， 及120有至少90%的同一性。 63. 如申請專利範圍第61項之經單離的核酸分子，其中該等 多核苷酸序列係各別與序列辨識編號：68、74、76、78、 80、82、84、86、88、90、92、94、96、98、100 ' 118， 及120有至少95%的同一性。 64. 如申請專利範圍第61項之經單離的核酸分子，其中該等 核酸分子各別包含序列辨識編號：68、74、76、78、80、 82、84、86、88、90、92、94、96、98、100、118，及 120的多核苷酸序列。 65. —種經單離的核酸分子，其係選自於以下所構成的群 組： (a) —核酸分子，其包含編碼一多肽之多核普酸序 列，其中該多肽包含與序列辨識編號：69有至少80%的 同一性之胺基酸序列，以及其中該多肽包含選自於以下 所構成的群組之PUFA合成酶活性：KS活性、MAT活性、 ACP活性、KR活性、DH活性、以及其等之組合； (b) —核酸分子，其包含編碼一多肽之多核苦酸序 16 201038734 列，其中該多肽包含與序列辨識編號：75有至少80%的 同一性之胺基酸序列，以及其中該多肽包含KS活性； (c) 一核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：77有至少80%的 同一性之胺基酸序列，以及其中該多肽包含MAT活性； (d) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：8卜83、85、87、 89、91、93、95、97或99的任何一者有至少80%的同一 性之胺基酸序列，以及其中該多肽包含ACP活性； (e) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：79有至少80%的 同一性之胺基酸序列，以及其中該多肽包含ACP活性； (f) 一核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：101有至少80% 的同一性之胺基酸序列，以及其中該多肽包含KR活 性；以及 (g) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：119有至少80% 的同一性之胺基酸序列，以及其中該多肽包含DH活性。 66. 如申請專利範圍第65項之經單離的核酸分子，其中該等 胺基酸序列各別與序列辨識編號：69、75、77、79、81、 83、85、87、89、91、93、95、97、99、101，以及 119 有至少90%的同一性。 67. 如申請專利範圍第65項之經單離的核酸分子，其中該等 17 201038734 胺基酸序列各別與序列辨識編號：69、75、77、79、81、 83、85、87、89、91、93、95、97、99、1(Π，以及 119 有至少95%的同一性。 68. 如申請專利範圍第65項之經單離的核酸分子，其中該等 多肽各別包含序列辨識編號·· 69、75、77、79、81、83、 85、87、89、91、93、95、97、99、101，以及 119的胺 基酸序列。 69. —種經單離的核酸分子，其係選自於以下所構成的群 組： (a) —核酸分子，其包含與序列辨識編號：70或序 列辨識編號：121有至少80%的同一性之多核苷酸序 列，其中該多核苷酸序列編碼一多肽，其包含選自於以 下所構成的群組之PUFA合成酶活性：KS活性、CLF活 性、AT活性、ER活性及其等之組合； (b) —核酸分子，其包含與序列辨識編號：102有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含KS活性； (c) 一核酸分子，其包含與序列辨識編號：104有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含CLF活性； (d) —核酸分子，其包含與序列辨識編號：106有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含AT活性；以及 (e) —核酸分子，其包含與序列辨識編號：108有至 18 201038734 y 80/。的同—性之多核芽酸序列，其中該多核苦酸序列 編碼一多肽，其包含ER活性e 70_如申μ專利關第69項之經單離的核酸分子，其中該等 夕核苦fee序列係、各別與序列辨識編號：⑽、1〇4、 1〇6、108，以及121有至少9〇%的同一性。 71·如申請專利範圍第69項之經單離的核酸分子，其中該等 多核替酸序列係各別與序列辨識編號：7〇、1〇2、刚、 1〇6、108,以及121有至少95%的同一性。 72·如申請專鄕圍第69項之轉離的核酸分子，其中該等 核酸分子各別包含序列辨識編號：7〇、ι〇2、ι〇4、1〇6、 108，以及121的多核苷酸序列。 73. -種經單離的核酸分子，其係選自於以下所構成的群 (a) -核酸分子’其包含編碼_多肽之多核普酸序 列，其中該多肽包含與序列辨識編號：71有至少8〇%的 同一性之胺基酸序列，以及其中該多肽包含選自於以下 所構成的群組之PUFA合成轉活性：ks活性、CLF活性、 AT活性、ER活性以及其等之組合； (b) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：103有至少80〇/〇 的同一性之胺基酸序列，以及其中該多肽包含KS活性； (c) 一核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：1〇5有至少8〇% 的同一性之胺基酸序列，以及其中該多肽包含CLF活 19 201038734 性； (d) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：107有至少80% 的同一性之胺基酸序列，以及其中該多肽包含AT活性； 以及 (e) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：109有至少80% 的同一性之胺基酸序列，以及其中該多肽包含ER活性。 74. 如申請專利範圍第73項之經單離的核酸分子，其中該等 胺基酸序列各別與序列辨識編號：71、103、105、107， 以及109有至少90%的同一性。 75. 如申請專利範圍第73項之經單離的核酸分子，其中該等 胺基酸序列各別與序列辨識編號：71、103、105、107， 以及109有至少95%的同一性。 76. 如申請專利範圍第73項之經單離的核酸分子，其中該等 多肽各別包含序列辨識編號：71、103、105、107，以 及109的胺基酸序列。 77. —種經單離的核酸分子，其係選自於以下所構成的群 組： (a) —核酸分子，其包含與序列辨識編號：72或序 列辨識編號：122有至少80%的同一性之多核苷酸序 列，其中該多核苷酸序列編碼一多肽，其包含選自於以 下所構成的群組之PUFA合成酶活性：DH活性、ER活性 及其等之組合； 20 201038734 (b) —核酸分子，其包含與序列辨識編號：11〇有至 少80%的同—性之多核倾序列其中該多料酸序列 編碼一多肽，其包含DH活性； (c) 一核酸分子，其包含與序列辨識編號：112有至 V 80/〇的同—性之多核苷酸序列，其中該多核苷酸序列 編碼—多肽，其包含DH活性；以及 (d) —核酸分子，其包含與序列辨識編號：114有至 少80%的同一性之多核苷酸序列，其中該多核苷酸序列 編碼一多肽，其包含ER活性。 78.如申請專利範圍第7 7項之經單離的核酸分子，其中該等 多核苷酸序列係各別與序列辨識編號：72、110、112、 114 ’以及122有至少90%的同一性。 79 ·如申請專利範圍第7 7項之經單離的核酸分子，其中該等 夕核芽酸序列係各別與序列辨識編號：72、110、112、 114 ’以及122有至少95%的同一性。 〇.如申請專利範圍第77項之經單離的核酸分子，其中該等 核酸分子各別包含序列辨識編號：72、110、112、114， 以及122的多核苷酸序列。 81_種經單離的核酸分子，其係選自於以下所構成的群 組： (a) —核酸分子，其包含編碼一多狀之多核苦酸序 列’其中該多肽包含與序列辨識編號：73有至少80%的 同—性之胺基酸序列，以及其中該多肽包含選自於以下 所構成的群組之PUFA合成酶活性：DH活性、er活性及 21 201038734 其等之組合； (b) -核酸分子’其包含編碼—多肽之多核普酸序 歹！其中6亥多肽包含與序列辨識編號：⑴有至少⑽％ 的同-性之胺錢序列，以及其中該多肽包含即活性； (c) 一核酸分子，其包含編碼一多肽之多核苷酸序 列’其中該多肽包含與序列辨識編號：113有至少贈。 的同-性之胺基酸序列，以及其中該多肽包含加活 性；以及 (d) —核酸分子，其包含編碼一多肽之多核苷酸序 列，其中該多肽包含與序列辨識編號：115有至少8〇% 的同一性之胺基酸序列，以及其中該多肽包#ER活性。 82.如申請專利範圍第81項之經單離的核酸分子其中該等 胺基酸序列各別與序列辨識編號：73、m、113，以及 115有至少90%的同一性。 83 如申請專利範圍第8丨項之經單離的核酸分子’其中該等 胺基酸序列各別與序列辨識編號·· 73、U1、113，以及 115有至少95%的同一性。 84·如申§青專利範圍第81項之經單離的核酸分子，其中該等 多肽各別包含序列辨識編號：73、111、113 ,及115之 胺基酸序列。 85. —種經單離的核酸分子，其包含編碼一多肽之多核苷酸 序列，該多肽包含選自於以下所構成的群組之PUFA合 成酶活性：KS活性、MAT活性、ACP活性、KR活性、 CLF活性、AT活性、ER活性、DH活性’以及其等之組 22 201038734 合， 其中該多核苷酸係於嚴苛條件下與申請專利範圍 第61-84項中任一項之多核苷酸序列之互補物雜交。 86. —種經單離的核酸分子，其包含一多核苷酸序列，其係 與申請專利範圍第61-84項中任一項之多核苷酸序列為 完全互補的。 87. —種重組型核酸分子，其包含如申請專利範圍第61-85 項中任一項之核酸分子或其等之組合以及一轉錄控制 〇 序列。 88. 如申請專利範圍第87項之重組型核酸分子，其中該重組 型核酸分子為一重組型載體。 、 89. —種宿主細胞，其表現如申請專利範圍第61-85項中任 -_ 一項之核酸分子、如申請專利範圍第87或88項之重組型 核酸分子，或其等之組合。 90. 如申請專利範圍第89項之宿主細胞，其中該宿主細胞係 選自於以下所構成的群組：植物細胞、微生物細胞，以 ❹ 及動物細胞。 91. 如申請專利範圍第90項之宿主細胞，其中該宿主細胞係 微生物細胞。 92. 如申請專利範圍第91項之微生物細胞，其中該微生物細 胞係一細菌。 93. 如申請專利範圍第91項之微生物細胞，其中該微生物細 胞為破囊壺菌。 94. 如申請專利範圍第93項之微生物細胞，其中該破囊壺菌 23 201038734 為裂殖壺菌屬或破囊壺菌屬。 95. —種產生至少一PUFA的方法，其包含： 於有效產生PUFA的條件下於一宿主細胞中表現 PUFA合成酶基因， 其中該PUFA合成酶基因包含如申請專利範圍第 61-85項中任一項之經單離的核酸分子、如申請專利範 圍第87或88項之重組型核酸分子，或其等之組合，以及 其中至少一個PUFA係被生產。 96 ·如申請專利範圍第9 5項之方法，其中該宿主細胞係選自 於以下所構成的群組：植物細胞、經單離的動物細胞， 以及微生物細胞。 97. 如申請專利範圍第95項之方法，其中該至少一個PUFA 包含DHA或二十碳五烯酸(EPA)。 98. —種生產富含DHA、EPA，或其等之組合之脂質的方 法，其包含： 於有效產生脂質的條件下於一宿主細胞中表現 PUFA合成酶基因，其中該PUFA合成酶基因包含如申請 專利範圍第61-85項中任一項之經單離的核酸分子、如 申請專利範圍第87或88項之重組型核酸分子，或其等之 組合於該宿主細胞中，以及 其中富含DHA、EPA，或其等之一組合的脂質係被 生產。 99. 一種製造重組型載體的方法，其包含將如申請專利範圍 第61-85項中任一項之經單離的核酸分子插入至一載體 24 201038734 内。 100. —種製造重組型宿主細胞的方法，其包含將如申請 專利範圍第99項之重組型載體引入至一宿主細胞中。 101. 如申請專利範圍第100項之方法，其中該宿主細胞係選 自於以下所構成的群組：植物細胞、經單離的動物細 胞，以及微生物細胞。 102•—種經單離的多肽，其係由申請專利範圍第61-85項中 任一項之多核苷酸序列所編碼。 103.—種經單離的多肽，其係選自於以下所構成的群組： (a) —多肽，其包含與序列辨識編號：69有至少80% 的同一性之胺基酸序列，其中該多肽包含選自於以下所 構成的群組之PUFA合成酶活性：KS活性、MAT活性、 ACP活性、KR活性、DH活性、以及其等之組合； (b) —多肽，其包含與序列辨識編號：75有至少80% 的同一性之胺基酸序列，其中該多肽包含KS活性； (c) 一多肽，其包含與序列辨識編號：77有至少80% 的同一性之胺基酸序列，其中該多肽包含MAT活性； (d) —多肽，其包含與序列辨識編號：81、83、85、 87、89、91、93、95、97或99的任一者有至少80%的同 一性之胺基酸序列，其中該多肽包含ACP活性； (e) —多肽，其包含與序列辨識編號：79有至少80% 的同一性之胺基酸序列，其中該多肽包含ACP活性； (f) 一多肽，其包含與序列辨識編號：101有至少 80%的同一性之胺基酸序列，其中該多肽包含KR活性； 25 201038734 以及 (g) —多肽，其包含與序列辨識編號：119有至少 80%的同一性之胺基酸序列，其中該多肽包含DH活性。 104. 如申請專利範圍第103項之經單離的多肽，其中該等胺 基酸序列各別與序列辨識編號：69、75、77、79、81、 83、85、87、89、91、93、95、97、99、101，以及 119 有至少90%的同一性。 105. 如申請專利範圍第103項之經單離的多肽，其中該等胺 基酸序列各別與序列辨識編號：69、75、77、79、81、 83、85、87、89、91、93、95、97、99、1(H，以及 119 有至少95%的同一性。 106. 如申請專利範圍第103項之經單離的多肽，其中該等多 肽各別包含序列辨識編號：69、75、77、79、81、83、 85、87、89、91、93、95、97、99、1(H，以及 119之胺 基酸序列。 107•—種經單離的多肽，其係選自於以下所構成的群組： (a) —多肽，其包含與序列辨識編號：71有至少80% 的同一性之胺基酸序列，其中該多肽包含選自於以下所 構成的群組之PUFA合成酶活性：KS活性、CLF活性、 AT活性、ER活性以及其等之組合； (b) —多肽，其包含與序列辨識編號：103有至少 80%的同一性之胺基酸序列，其中該多肽包含KS活性； (c) 一多肽，其包含與序列辨識編號：105有至少 80%的同一性之胺基酸序列，其中該多肽包含CLF活性； 26 201038734 (d) —多肽，其包含與序列辨識編號：107有至少 80%的同一性之胺基酸序列，其中該多肽包含AT活性； 以及 (e) —多肽，其包含與序列辨識編號：109有至少 80%的同一性之胺基酸序列，其中該多肽包含ER活性。 108. 如申請專利範圍第107項之經單離的多肽，其中該等胺 基酸序列各別與序列辨識編號：71、103、105、107， 以及109有至少90%的同一性。 109. 如申請專利範圍第107項之經單離的多肽，其中該等胺 基酸序列各別與序列辨識編號：71、103、105、107， 以及109有至少95%的同一性。 110. 如申請專利範圍第107項之經單離的多肽，其中該等多 肽各別包含序列辨識編號：71、103、105、107，以及 109的胺基酸序列。 111. 一種經單離的多肽，其係選自於以下所構成的群組： (a) —多肽，其包含與序列辨識編號：73有至少80% 的同一性之胺基酸序列，其中該多肽包含選自於以下所 構成的群組之PUFA合成酶活性：DH活性、ER活性以及 其等之組合； (b) —多肽，其包含與序列辨識編號：111有至少 80%的同一性之胺基酸序列，其中該多肽包含DH活性； (c) 一多肽，其包含與序列辨識編號：113有至少 80%的同一性之胺基酸序列，其中該多肽包含DH活性； 以及 27 201038734 (d) —多肽，其包含與序列辨識編號：115有至少 80%的同一性之胺基酸序列，其中該多肽包含ER活性。 112. 如申請專利範圍第111項之經單離的多肽，其中該等胺 基酸序列各別與序列辨識編號：73、111、113，以及115 有至少90%的同一性。 113. 如申請專利範圍第ui項之經單離的多肽，其中該等胺 基酸序列各別與序列辨識編號：73、111、113，以及115 有至少95%的同一性。 114. 如申請專利範圍第111項之經單離的多肽，其中該等多 肽各別包含序列辨識編號：73、111、113，以及115之 胺基酸序列。 115. 如申請專利範圍第102-114項中任一項之經單離的多 肽，其中該多肽為一融合多肽。 116. —種組成物，其包含如申請專利範圍第i〇2_U4項中任 一項之多肽以及生物上可接受的載體。 Π7.—種於具有PUFA合成酶活性的有機體内增加〇ΗΑ、 EPA，或其等之一組合的生產之方法，其包含： 於有效產生DHA、EPA或其等之一組合的條件下於 該有機體中表現如申請專利範圍第61_85項中任一項之 經單離的核酸分子、如申請專利範圍第87或88項之重組 型核酸分子，或其等之組合， 其中該PUFA合成酶活性於該有機體中取代不活化 或被刪除的活性、引入新的活性、或提高現存的活性， 以及 28 201038734 其中該有機體中的DHA、EPA，或其等之一組合之 生產為增加的。 118. —種自宿主細胞單離脂質的方法，其包含： (a)於有效產生脂質的條件下於該宿主細胞中表現 PUFA合成酶基因，其中PKS系統包含如申請專利範圍第 61-85項中任一項之經單離的核酸分子、如申請專利範 圍第87或88項之重組型核酸分子，或其等之組合於該宿 主細胞中，以及 〇 ^ (b)自該宿主細胞單離脂質。 119. 如申請專利範圍第118項之方法，其中該宿主細胞係選 自於以下所構成的群組：植物細胞、經單離的動物細 ' - 胞，以及微生物細胞。 _ _ 120.如申請專利範圍第118項之方法，其中該等脂質包含 DHA、EPA，或其等之一組合。 〇 29(d) - the nucleus, wherein any of the plurality or 24 has at least 80% of the same (g)-nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide wherein the polypeptide comprises and the sequence identification number: 28 has at least 80% identity amino acid sequence, and wherein the polypeptide comprises DH activity. 6_ The isolated nucleic acid molecule of claim 5, wherein the amino acid sequence and the sequence identification number are: 2, 8, 1, 12, 14, 16, 16, 18, 22, 24 , 26, and 28 each have at least 90% identity. 7. An isolated nucleic acid molecule according to claim 5, wherein the amino acid sequence and the sequence identification number are: 2, 8, 1, 12, 14, 16, 18, 20, 22, 24, 26, And 28 have at least 95% identity. 3 201038734 8.  An isolated nucleic acid molecule according to claim 5, wherein the polypeptides each comprise a sequence identification number: 2, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, and 28 Amino acid sequence 歹 ij ° 9.  An isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 3, wherein The polynucleotide sequence encodes a polypeptide comprising PUFA synthase activity selected from the group consisting of KS activity, chain length factor (CLF) activity, thiol transferase (AT) activity, olefinic group- ACP reductase (ER) activity, and combinations thereof; (b) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 29, wherein the polynucleotide sequence encodes a a polypeptide comprising KS activity; (c) a nucleic acid molecule comprising a polynucleotide sequence at least 80% identical to SEQ ID NO: 31, wherein the polynucleotide sequence encodes a polypeptide comprising CLF (d) - a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 33, wherein the polynucleotide sequence encodes a polypeptide comprising AT activity; and (e) - a nucleic acid molecule comprising a sequence Identification number: 35 polynucleotide sequence identity to at least 80%, wherein the polynucleotide sequence encoding a polypeptide comprising ER activity. 10.  An isolated nucleic acid molecule according to claim 9, wherein the 4 201038734 polynucleotide sequences are at least 90% identical to the sequence identification numbers: 3, 29, 31, 33, and 35, respectively. 11.  An isolated nucleic acid molecule according to claim 9 wherein the polynucleotide sequences are at least 95% identical to the sequence identification numbers: 3, 29, 31, 33, and 35, respectively. 12.  The isolated nucleic acid molecule of claim 9, wherein the nucleic acid molecules each comprise a polynucleotide sequence of sequence number: 3, 29, 31, 33, and 35. 13.  An isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises and the sequence identification number: 4 An amino acid sequence having at least 80% identity, and wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of KS activity, CLF activity, AT activity, ER activity, and the like (b) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 30, and wherein the polypeptide comprises KS activity (c) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence number: 32, and wherein the polypeptide comprises CLF activity (d) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence number: 34, and wherein the polypeptide comprises an AT And 5, 201038734 (e) - a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 36, and wherein The polypeptide contains ER activity. 14.  An isolated nucleic acid molecule according to claim 13 wherein the amino acid sequences are at least 90% identical to the sequence identification numbers: 4, 30, 32, 34, and 36, respectively. 15.  An isolated nucleic acid molecule according to claim 13 wherein the amino acid sequences are at least 95% identical to the sequence identification numbers: 4, 30, 32, 34, and 36, respectively. 16.  The isolated nucleic acid molecule of claim 13, wherein the polypeptides each comprise an amino acid sequence of sequence identification numbers: 4, 30, 32, 34, and 36. 17.  An isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 5, wherein The polynucleotide sequence encodes a polypeptide comprising a PUFA synthase activity selected from the group consisting of DH activity, ER activity, and combinations thereof; (b) - a nucleic acid molecule comprising and sequence Identification number: 37 has a polynucleotide sequence of at least 80% identity, wherein the polynucleotide sequence encodes a polypeptide comprising DH activity; (c) a nucleic acid molecule comprising at least a sequence identification number: 39 80% identity polynucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising DH activity; and 201038734 (d) - a nucleic acid molecule comprising at least 80% identical to the sequence ID: 41 A polynucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising ER activity. 18.  An isolated nucleic acid molecule according to claim 17, wherein the polynucleotide sequences are at least 90% identical to the sequence identification numbers: 5, 37, 39, and 41, respectively. 19.  An isolated nucleic acid molecule according to claim 17, wherein the polynucleotide sequences are at least 95% identical to the sequence identification numbers: 5, 37, 39, and 41, respectively. 20.  An isolated nucleic acid molecule according to claim 17, wherein the nucleic acid molecules each comprise a polynucleotide sequence of sequence identification numbers: 5, 37, 39, and 41. twenty one.  An isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises and the sequence identification number: 6 An amino acid sequence having at least 80% identity, and wherein the polypeptide comprises a combination of PUFA synthase activity, DH activity, ER activity, and the like selected from the group consisting of: (b) - a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence number: 38, and wherein the polypeptide comprises DH activity; (c) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% of the 7 201038734 identity with sequence number: 40, and wherein the polypeptide comprises DH activity; d) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 42, and wherein the polypeptide comprises ER Sex. twenty two.  An isolated nucleic acid molecule according to claim 21, wherein the amino acid sequences are at least 90% identical to the sequence identification numbers: 6, 38, 40, and 42. twenty three.  An isolated nucleic acid molecule according to claim 21, wherein the amino acid sequences are at least 95% identical to the sequence identification numbers: 6, 38, 40, and 42. twenty four.  The isolated nucleic acid molecule of claim 21, wherein the polypeptides each comprise an amino acid sequence of sequence identification numbers: 6, 38, 40, and 42. 25.  An isolated nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide comprising PUFA synthase activity selected from the group consisting of KS activity, MAT activity, ACP activity, KR activity a combination of CLF activity, AT activity, ER activity, DH activity, and the like, wherein the polynucleotide is complementary to the polynucleotide sequence of any one of claims 1 to 24 under severe conditions. Hybridization. 26.  An isolated nucleic acid molecule comprising a polynucleotide sequence which is fully complementary to the polynucleotide sequence of any one of claims 1 to 24. 201038734 27.  A recombinant nucleic acid molecule comprising a nucleic acid molecule according to any one of claims 1 to 25, or a combination thereof, and a transcription control sequence. 28.  A recombinant nucleic acid molecule according to claim 27, wherein the recombinant nucleic acid molecule is a recombinant vector. 29.  A host cell which exhibits a nucleic acid molecule according to any one of claims 1 to 25, a recombinant nucleic acid molecule as claimed in claim 27 or 28, or a combination thereof. 30.  The host cell of claim 29, wherein the host cell line is selected from the group consisting of a plant cell, a microbial cell, and an animal cell. 31.  The host cell of claim 30, wherein the host cell is a microbial cell. 32.  A microbial cell according to claim 31, wherein the microbial cell is a bacterium. 33.  The microbial cell of claim 31, wherein the microbial cell is a Thraustochytrium. 34.  The microbial cell of claim 33, wherein the Thraustochytrium is Schizochytrium or Thraustochytrium.  A method of producing at least one PUFA comprising: expressing a PUFA synthase gene in a host cell under conditions effective to produce a PUFA, wherein the PUFA synthase gene comprises, as claimed in claim 9, 2010.  37.  38.  39.  40.  41.  42.   An isolated nucleic acid molecule according to any one of items 1 to 25, a recombinant nucleic acid molecule according to claim 27 or 28, or a combination thereof, and at least one of which is produced. The method of claim 35, wherein the host cell line is selected from the group consisting of a plant cell, an isolated animal cell, and a microbial cell. The method of claim 35, wherein the at least one PIJFA comprises docosahexaenoic acid (DHA). A method for producing a DHA-rich lipid, comprising: expressing a PUFA synthase gene in a host cell under conditions effective to produce a lipid, wherein the PUFA synthase gene comprises any one of claims 1 to 25 An isolated nucleic acid molecule, a recombinant nucleic acid molecule as claimed in claim 27 or 28, or a combination thereof in the host cell, and a DHA-rich lipid thereof are produced. A method of producing a recombinant vector comprising the insertion of a ligated nucleic acid molecule according to any one of claims 1 to 25 into a carrier. A method of producing a recombinant plastic host cell comprising introducing a recombinant vector according to claim 39 of the patent application into a host cell. The method of claim 4, wherein the host cell line is selected from the group consisting of a plant cell, an isolated animal cell, and a microbial cell. An isolated polypeptide which is encoded by the polynucleotide sequence of any of 201038734 in the scope of the patent application. 43.  An isolated polypeptide which is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 2, wherein the polypeptide A PUFA synthase activity comprising a group selected from the group consisting of KS activity, MAT activity, ACP activity, KR activity, DH activity, and combinations thereof; (b) - a polypeptide comprising the sequence identification number: 8 having at least 80% identity amino acid sequence, wherein the polypeptide comprises KS activity; (c) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence number: 10, wherein The polypeptide comprises MAT activity; (d) a polypeptide comprising an amino acid sequence having at least 80% identity to any one of sequence identification number: 14, 16, 18, 20, 22, or 24, wherein The polypeptide comprises ACP activity; (e) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 12, wherein the polypeptide comprises ACP activity; (f) a polypeptide comprising the sequence Identification number: 26 amino acid sequence having at least 80% identity, The polypeptide comprising a KR activity; and (g) - a polypeptide comprising a sequence identification number: 28, at least 80% identity to the amino acid sequence, wherein the polypeptide comprises DH activity. 44.  An isolated polypeptide according to claim 43, wherein the amino acid sequences are each different from the sequence identification numbers: 2, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, And 28 have at least 90% identity. 45.  The isolated polypeptide of claim 43, wherein the amino acid 11 201038734 acid sequence and the sequence identification number: 2, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, and 28 have at least 95% identity. 46.  The isolated polypeptide of claim 43, wherein the polypeptides each comprise a sequence identification number: 2, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, and 28 Amino acid sequence. 47.  An isolated polypeptide which is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 4, wherein the polypeptide A PUFA synthase activity comprising a group selected from the group consisting of KS activity, CLF activity, AT activity, ER activity, and combinations thereof; (b) a polypeptide comprising at least a sequence identification number: 30 80% identical amino acid sequence, wherein the polypeptide comprises KS activity; (c) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 32, wherein the polypeptide comprises CLF activity; (d) - a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 34, wherein the polypeptide comprises AT activity; and (e) - a polypeptide comprising a sequence identification number :36 an amino acid sequence having at least 80% identity, wherein the polypeptide comprises ER activity. 48.  An isolated polypeptide according to claim 47, wherein the amino acid sequences are each at least 90°/SEQ ID NO: 4, 30, 32, 34, and 36. Identity. 49.  The isolated polypeptide of claim 47, wherein the amino acid sequences are at least 95% identical to the sequence identification numbers: 4, 30, 32, 34, and 36, respectively. 12 201038734 50.  The isolated polypeptide of claim 47, wherein the polypeptides each comprise an amino acid sequence of sequence identification numbers: 4, 30, 32, 34, and 36. 51.  An isolated polypeptide which is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 6, wherein the polypeptide A PUFA synthase activity comprising a group selected from the group consisting of DH activity, ER activity, and combinations thereof; ^ (b) - a polypeptide comprising at least 80% identity to sequence identification number: 38 An amino acid sequence, wherein the polypeptide comprises DH activity; (c) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence number: 40, wherein the polypeptide comprises DH activity; - - and (d) - a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 42, wherein the polypeptide comprises ER activity. 52.  An isolated polypeptide according to claim 51, wherein the amino acid sequences are at least 90% identical to the sequence identification numbers: 6, 38, 40, and 42. 53.  The isolated polypeptide of claim 51, wherein the amino acid sequences are at least 95% identical to the sequence identification numbers: 6, 38, 40, and 42. 54.  The isolated polypeptide of claim 51, wherein the polypeptides each comprise an amino acid sequence of sequence identification numbers: 6, 38, 40, and 42. 13 201038734 55.  The isolated polypeptide of any one of claims 42-54, wherein the polypeptide is a fusion polypeptide. 56.  A composition comprising a polypeptide according to any one of claims 42-54 and a biologically acceptable carrier. 57.  - A method for increasing production of DHA in an organism having PUFA synthase activity, comprising: expressing in the organism under conditions effective to produce DHA, as shown in any one of claims 1 to 25 A nucleic acid molecule, such as the recombinant nucleic acid molecule of claim 27 or 28, or a combination thereof, wherein the PUFA synthase activity replaces an activity that is not activated or deleted in the organism, introduces a new activity, or Increasing the existing activity, and wherein the production of DHA in the organism is increased. 58.  - A method for detaching a lipid from a host cell, comprising: (a) expressing a PUFA synthase gene in the host cell under conditions effective to produce a lipid, wherein the PKS system comprises any of claims 1-25 An isolated nucleic acid molecule, such as the recombinant nucleic acid molecule of claim 27 or 28, or a combination thereof in the host cell, and (b) the isolated lipid from the host cell. 59.  The method of claim 58, wherein the host cell line is selected from the group consisting of plant cells, isolated animal cells, and microbial cells. 14 201038734 60.  The method of claim 58, wherein the lipids comprise DHA. 61.  An isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising at least 80% identity to sequence identification number: 68 or sequence identification number: 120 a polynucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising PUFA synthase activity selected from the group consisting of KS activity, MAT activity, ACP activity, KR activity, DH activity, and a combination of: (b) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 74, wherein the polynucleotide sequence encodes a polypeptide comprising KS activity; a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 76, wherein the polynucleotide sequence encodes a polypeptide comprising MAT activity; (d) a nucleic acid molecule A polynucleotide sequence comprising at least 80% identity to any one of sequence identification numbers: 80, 82, 84, 86, 88, 90, 92, 94, 96 or 98, wherein the polynucleotide sequence encodes a polynucleotide sequence a polypeptide comprising ACP activity; (e) - An acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 78, wherein the polynucleotide sequence encodes a polypeptide comprising ACP activity; (f) a nucleic acid molecule comprising Sequence identification number: 100 has a polynucleotide sequence of at least 80% identity, wherein the polynucleotide sequence 15 201038734 encodes a polypeptide comprising KR activity; and (g) a nucleic acid molecule comprising the sequence identification number : 118 A polynucleotide sequence having at least 80% identity, wherein the polynucleotide sequence encodes a polypeptide comprising DH activity. 62.  An isolated nucleic acid molecule according to claim 61, wherein the polynucleotide sequences are individually and sequence identification numbers: 68, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 118, and 120 have at least 90% identity. 63.  An isolated nucleic acid molecule according to claim 61, wherein the polynucleotide sequences are individually and sequence identification numbers: 68, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100 '118, and 120 have at least 95% identity. 64.  An isolated nucleic acid molecule according to claim 61, wherein the nucleic acid molecules each comprise a sequence identification number: 68, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94 , 96, 98, 100, 118, and 120 polynucleotide sequences. 65.  An isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises and the sequence identification number: 69 An amino acid sequence having at least 80% identity, and wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of KS activity, MAT activity, ACP activity, KR activity, DH activity, and a combination of: (b) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide of 16 201038734, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 75, and Wherein the polypeptide comprises KS activity; (c) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 77, and Wherein the polypeptide comprises MAT activity; (d) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises and the sequence identification number: 8, 83, 85, 87, 89, 91, 93, 95 , 97 or 99 Any one of which has at least 80% identity amino acid sequence, and wherein the polypeptide comprises ACP activity; (e) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises a sequence Identification number: 79 amino acid sequence having at least 80% identity, and wherein the polypeptide comprises ACP activity; (f) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises a sequence Identification number: 101 having at least 80% identity amino acid sequence, and wherein the polypeptide comprises KR activity; and (g) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises Sequence Identification Number: 119 An amino acid sequence having at least 80% identity, and wherein the polypeptide comprises DH activity. 66.  An isolated nucleic acid molecule according to claim 65, wherein the amino acid sequences are individually associated with sequence identification numbers: 69, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93 , 95, 97, 99, 101, and 119 have at least 90% identity. 67.  An isolated nucleic acid molecule according to claim 65, wherein the 17 201038734 amino acid sequence and the sequence identification number: 69, 75, 77, 79, 81, 83, 85, 87, 89, 91 , 93, 95, 97, 99, 1 (Π, and 119 have at least 95% identity. 68.  An isolated nucleic acid molecule according to claim 65, wherein the polypeptides each comprise a sequence identification number 69, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95 The amino acid sequences of 97, 99, 101, and 119. 69.  An isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising at least 80% identity to sequence identification number: 70 or sequence identification number: 121 a polynucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising a PUFA synthase activity selected from the group consisting of KS activity, CLF activity, AT activity, ER activity, and the like; (b) a nucleic acid molecule comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 102, wherein the polynucleotide sequence encodes a polypeptide comprising KS activity; (c) a nucleic acid molecule , comprising a polynucleotide sequence having at least 80% identity to sequence identification number: 104, wherein the polynucleotide sequence encodes a polypeptide comprising CLF activity; (d) - a nucleic acid molecule comprising: sequence identification ID: 106 has a polynucleotide sequence of at least 80% identity, wherein the polynucleotide sequence encodes a polypeptide comprising AT activity; and (e) a nucleic acid molecule comprising: and sequence identification number: 108 18 2010387 34 y 80/. a polymorphic nucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising an ER activity e 70 - such as an isolated nucleic acid molecule of claim 69, wherein the The fee sequence, the individual and the sequence identification number: (10), 1〇4, 1〇6, 108, and 121 have at least 9% identity. 71. The isolated nucleic acid molecule of claim 69, wherein the polynuclear acid sequence is different from the sequence identification number: 7〇, 1〇2, just, 1〇6, 108, and 121 At least 95% identity. 72. If you apply for a nucleic acid molecule that is specifically transferred to the 69th item, wherein the nucleic acid molecules each comprise a sequence identification number: 7〇, ι〇2, ι〇4, 1〇6, 108, and 121 Glycosidic acid sequence. 73.  An isolated nucleic acid molecule selected from the group consisting of: a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least 8 with a sequence number: 71 An amino acid sequence of 〇% identity, and wherein the polypeptide comprises a PUFA synthesis transactivation activity selected from the group consisting of: ks activity, CLF activity, AT activity, ER activity, and combinations thereof; a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80 〇/〇 identity to Sequence ID: 103, and wherein the polypeptide comprises KS activity; (c) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 8% identity with a sequence number: 1〇5, and wherein the polypeptide comprises CLF live 19 201038734; (d) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to sequence identification number: 107, and The polypeptide comprises AT activity; and (e) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 80% identity to Sequence ID: 109, and Wherein the polypeptide comprises ER activity. 74.  An isolated nucleic acid molecule according to claim 73, wherein the amino acid sequences are each at least 90% identical to the sequence identification numbers: 71, 103, 105, 107, and 109. 75.  An isolated nucleic acid molecule according to claim 73, wherein the amino acid sequences are each at least 95% identical to the sequence identification numbers: 71, 103, 105, 107, and 109. 76.  An isolated nucleic acid molecule according to claim 73, wherein the polypeptides each comprise a sequence identification number: 71, 103, 105, 107, and an amino acid sequence of 109. 77.  An isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising at least 80% identity to sequence identification number: 72 or sequence identification number: 122 A polynucleotide sequence, wherein the polynucleotide sequence encodes a polypeptide comprising a PUFA synthase activity selected from the group consisting of DH activity, ER activity, and combinations thereof; 20 201038734 (b) - a nucleic acid molecule comprising a multi-nuclear decantation sequence having at least 80% homology to a sequence identification number: 11〇, wherein the polyacid sequence encodes a polypeptide comprising DH activity; (c) a nucleic acid molecule comprising Sequence Identification Number: 112 has a homologous polynucleotide sequence to V80/〇, wherein the polynucleotide sequence encodes a polypeptide comprising DH activity; and (d) a nucleic acid molecule comprising a sequence identification number : 114 A polynucleotide sequence having at least 80% identity, wherein the polynucleotide sequence encodes a polypeptide comprising ER activity. 78. An isolated nucleic acid molecule according to claim 7 wherein the polynucleotide sequences are at least 90% identical to the sequence identification numbers: 72, 110, 112, 114' and 122, respectively. 79. The isolated nucleic acid molecule of claim 7, wherein the nucleotide sequences are at least 95% identical to the sequence identification numbers: 72, 110, 112, 114' and 122. Sex. Hey. An isolated nucleic acid molecule according to claim 77, wherein the nucleic acid molecules each comprise a polynucleotide sequence of sequence identification numbers: 72, 110, 112, 114, and 122. An isolated nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule comprising a polynucleic acid sequence encoding a polymorphism wherein the polypeptide comprises and the sequence identification number: 73 having at least 80% homo-amino acid sequence, and wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of DH activity, er activity, and 21 201038734; ) - a nucleic acid molecule that contains a multi-nucleotide acid sequence encoding a polypeptide! Wherein the 6H polypeptide comprises a sequence number of (1) having at least (10)% homologous amine money, and wherein the polypeptide comprises an activity; (c) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide 'The polypeptide contains at least a gift with sequence identification number: 113. a homo-amino acid sequence, wherein the polypeptide comprises an additional activity; and (d) a nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises at least one of the sequence identification number: 115 8 〇 % of the identity of the amino acid sequence, and wherein the polypeptide comprises #ER activity. 82. An isolated nucleic acid molecule according to claim 81, wherein the amino acid sequences are each at least 90% identical to the sequence identification numbers: 73, m, 113, and 115. 83. The isolated nucleic acid molecule&apos; as claimed in claim 8 wherein each of the amino acid sequences is at least 95% identical to the sequence identification number 73, U1, 113, and 115. 84. The isolated nucleic acid molecule of claim 81, wherein the polypeptides each comprise an amino acid sequence of sequence identification numbers: 73, 111, 113, and 115. 85.  An isolated nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide comprising PUFA synthase activity selected from the group consisting of KS activity, MAT activity, ACP activity, KR activity , CLF activity, AT activity, ER activity, DH activity 'and its group 22 201038734, wherein the polynucleotide is a polynucleotide under severe conditions and any one of claims 61-84. The complement of the sequence is hybridized. 86.  An isolated nucleic acid molecule comprising a polynucleotide sequence which is fully complementary to the polynucleotide sequence of any one of claims 61-84. 87.  A recombinant nucleic acid molecule comprising a nucleic acid molecule according to any one of claims 61-85, or a combination thereof, and a transcription control 〇 sequence. 88.  A recombinant nucleic acid molecule according to claim 87, wherein the recombinant nucleic acid molecule is a recombinant vector. 89.  A host cell which exhibits a nucleic acid molecule according to any one of claims 61-85, a recombinant nucleic acid molecule according to claim 87 or 88, or a combination thereof. 90.  The host cell of claim 89, wherein the host cell line is selected from the group consisting of a plant cell, a microbial cell, and an animal cell. 91.  The host cell of claim 90, wherein the host cell is a microbial cell. 92.  A microbial cell as claimed in claim 91, wherein the microbial cell is a bacterium. 93.  The microbial cell of claim 91, wherein the microbial cell is a Thraustochytrium. 94.  The microbial cell of claim 93, wherein the Thraustochytrium 23 201038734 is a Schizochytrium or a Thraustochytrium. 95.  A method for producing at least one PUFA, comprising: expressing a PUFA synthase gene in a host cell under conditions effective to produce a PUFA, wherein the PUFA synthase gene comprises any one of claims 61-85 An isolated nucleic acid molecule, such as a recombinant nucleic acid molecule of claim 87 or 88, or a combination thereof, and at least one of which is produced. 96. The method of claim 9, wherein the host cell line is selected from the group consisting of plant cells, isolated animal cells, and microbial cells. 97.  The method of claim 95, wherein the at least one PUFA comprises DHA or eicosapentaenoic acid (EPA). 98.  A method for producing a lipid rich in DHA, EPA, or a combination thereof, comprising: expressing a PUFA synthase gene in a host cell under conditions effective for producing a lipid, wherein the PUFA synthase gene comprises a patent application The isolated nucleic acid molecule of any one of the items 61-85, the recombinant nucleic acid molecule of claim 87 or 88, or a combination thereof in the host cell, and wherein the DHA is enriched, A lipid system of EPA, or a combination thereof, is produced. 99.  A method of producing a recombinant vector comprising inserting the isolated nucleic acid molecule according to any one of claims 61-85 into a vector 24 201038734. 100.  A method of producing a recombinant host cell comprising introducing a recombinant vector according to claim 99 of the patent application into a host cell. 101.  The method of claim 100, wherein the host cell line is selected from the group consisting of plant cells, isolated animal cells, and microbial cells. 102. An isolated polypeptide which is encoded by the polynucleotide sequence of any one of claims 61-85. 103. An isolated polypeptide which is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 69, wherein the polypeptide A PUFA synthase activity comprising a group selected from the group consisting of KS activity, MAT activity, ACP activity, KR activity, DH activity, and combinations thereof; (b) - a polypeptide comprising: and a sequence identification number: 75 having at least 80% identity amino acid sequence, wherein the polypeptide comprises KS activity; (c) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 77, wherein The polypeptide comprises MAT activity; (d) - a polypeptide comprising at least 80% identity to any one of sequence identification numbers: 81, 83, 85, 87, 89, 91, 93, 95, 97 or 99 An amino acid sequence, wherein the polypeptide comprises ACP activity; (e) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 79, wherein the polypeptide comprises ACP activity; (f) a polypeptide comprising an amine having at least 80% identity to sequence identification number: 101 A base acid sequence, wherein the polypeptide comprises KR activity; 25 201038734 and (g) - a polypeptide comprising an amino acid sequence having at least 80% identity to SEQ ID NO: 119, wherein the polypeptide comprises DH activity. 104.  The isolated polypeptide of claim 103, wherein the amino acid sequences are different from the sequence identification numbers: 69, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, and 119 have at least 90% identity. 105.  The isolated polypeptide of claim 103, wherein the amino acid sequences are different from the sequence identification numbers: 69, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 1 (H, and 119 have at least 95% identity. 106.  An isolated polypeptide according to claim 103, wherein the polypeptides each comprise a sequence identification number: 69, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97 , 99, 1 (H, and 119 amino acid sequence. 107 • an isolated polypeptide, which is selected from the group consisting of: (a) - a polypeptide, which contains and the sequence identification number: 71 having at least 80% identity amino acid sequence, wherein the polypeptide comprises PUFA synthase activity selected from the group consisting of KS activity, CLF activity, AT activity, ER activity, and combinations thereof; (b) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 103, wherein the polypeptide comprises KS activity; (c) a polypeptide comprising: and a sequence identification number: 105 At least 80% identical amino acid sequence, wherein the polypeptide comprises CLF activity; 26 201038734 (d) - a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 107, wherein The polypeptide comprises AT activity; and (e) - the polypeptide, which comprises and recognizes the sequence ID: 109 has at least 80% identity amino acid sequence, wherein the polypeptide comprises ER activity.  An isolated polypeptide according to claim 107, wherein the amino acid sequences are each at least 90% identical to the sequence identification numbers: 71, 103, 105, 107, and 109. 109.  An isolated polypeptide according to claim 107, wherein the amino acid sequences are each at least 95% identical to the sequence identification numbers: 71, 103, 105, 107, and 109. 110.  An isolated polypeptide according to claim 107, wherein the polypeptides each comprise an amino acid sequence of sequence identification numbers: 71, 103, 105, 107, and 109. 111.  An isolated polypeptide which is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 73, wherein the polypeptide comprises A PUFA synthase activity selected from the group consisting of DH activity, ER activity, and combinations thereof; (b) a polypeptide comprising an amino group having at least 80% identity to sequence identification number: 111 An acid sequence, wherein the polypeptide comprises DH activity; (c) a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 113, wherein the polypeptide comprises DH activity; and 27 201038734 (d) - a polypeptide comprising an amino acid sequence having at least 80% identity to sequence identification number: 115, wherein the polypeptide comprises ER activity. 112.  The isolated polypeptide of claim 111, wherein the amino acid sequences are each at least 90% identical to the sequence identification numbers: 73, 111, 113, and 115. 113.  The isolated polypeptide of claim ui, wherein the amino acid sequences are at least 95% identical to the sequence identification numbers: 73, 111, 113, and 115, respectively. 114.  The isolated polypeptide of claim 111, wherein the polypeptides each comprise an amino acid sequence of sequence identification numbers: 73, 111, 113, and 115. 115.  The isolated polypeptide of any one of claims 102-114, wherein the polypeptide is a fusion polypeptide. 116.  A composition comprising a polypeptide according to any one of claims 2 to 2, and a biologically acceptable carrier. Π7. - a method of producing a combination of hydrazine, EPA, or the like in an organism having PUFA synthetase activity, comprising: in the organism under conditions effective to produce a combination of DHA, EPA, or the like An isolated nucleic acid molecule according to any one of claims 61 to 85, wherein the recombinant nucleic acid molecule of claim 87 or 88, or a combination thereof, wherein the PUFA synthase is active in the organism Substituting for activity that is not activated or deleted, introducing new activity, or increasing existing activity, and 28 201038734 wherein the production of a combination of DHA, EPA, or the like in the organism is increased. 118.  A method for detaching a lipid from a host cell, comprising: (a) expressing a PUFA synthase gene in the host cell under conditions effective to produce a lipid, wherein the PKS system comprises any of those in the scope of claims 61-85 An isolated nucleic acid molecule, such as the recombinant nucleic acid molecule of claim 87 or 88, or a combination thereof, and (b) isolated from the host cell. 119.  The method of claim 118, wherein the host cell line is selected from the group consisting of a plant cell, an isolated animal cell, and a microbial cell. _ _ 120. The method of claim 118, wherein the lipid comprises DHA, EPA, or a combination thereof. 〇 29
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